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PCR Enzymes TransFastTaqDNA Polymerase

EasyTaq

DNA Polymerase

EasyTaq

DNA Polymerase for PAGE

TransTaq-T DNA Polymerase

TransTaq

DNA Polymerase High Fidelity (HiFi)

TransStartTaqDNA Polymerase

TransStartTopTaqDNA Polymerase

EasyPfuDNA Polymerase TransStart

FastPfuDNA Polymerase

TransStartFastPfuFly DNA Polymerase

GC Enhancer

PCR Stimulant

PCR SuperMix

2EasyTaq

PCR SuperMix

2EasyTaqPCR SuperMix for PAGE

2TransTaq-T PCR SuperMix

2TransTaq High Fidelity (HiFi) PCR SuperMix

2EasyPfuPCR SuperMix

2TransStartFastPfuPCR SuperMix

Direct PCR

TransDirectTM

Animal Tissue PCR Kit

TransDirectTM Plant Tissue PCR Kit

TransDirectTM

Blood PCR Kit

RT-PCR

EasyScript Reverse Transcriptase

TransScript

Reverse Transcriptase

TransScript II Reverse Transcriptase

EasyScript

First-Strand cDNA Synthesis SuperMix

TransScript First-Strand cDNA Synthesis SuperMix

TransScript

One-Step gDNA Removal and cDNA

Synthesis SuperMix

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006

007

009

010

012

014

016017

019

021

022

024

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026

Chapter 1 PCR, RT-PCR,qPCR and qRT-PCR

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045


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qPCR and qRT-PCR SuperMix

TransStart

Green qPCR SuperMix

TransStart Green qPCR SuperMix UDG

TransStart

Top Green qPCR SuperMix

TransScript

Green Two-Step qRT-PCR SuperMix

TransScript

II Green Two-Step qRT-PCR SuperMix

TransScript

Green One-Step qRT-PCR SuperMix

TransScript II Green One-Step qRT-PCR SuperMix

TransStart

Probe qPCR SuperMix

TransScript Probe One-Step qRT-PCR SuperMix TransScript

II Probe One-Step qRT-PCR SuperMix

High Purity dNTPs

High Purity dNTPs

Chapter 1 PCR RT-PCR qPCR and qRT-PCR

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060

062

063

064

066

068

070

072

073

TransScript

All-in-One First-Strand cDNA Synthesis

SuperMix for PCR

TransScript

All-in-One First-Strand cDNA Synthesis

SuperMix for qPCR

TransScriptII First-Strand cDNA Synthesis SuperMix

TransScript

II One-Step gDNA Removal and cDNA

Synthesis SuperMix

TransScript

II All-in-One First-Strand cDNA Synthesis

SuperMix for PCR

TransScript II All-in-One First-Strand cDNA SynthesisSuperMix for qPCR

TransScript

Two-Step RT-PCR SuperMix

TransScriptII Two-Step RT-PCR SuperMix

EasyScript One-Step RT-PCR SuperMix

TransScript

One-Step RT-PCR SuperMix

TransScript II One-Step RT-PCR SuperMix

Ribonuclease Inhibitor

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PCR Enzymes Chapter 1

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PCR EnzymesPCR enzymes are puried from E.coli strains carrying genes for specic DNA polymerase. The choice of DNA polymerase

for PCR application is highly dependent on the characteristics of the system as well as the desired results. The following is

PCR Enzyme Selection Guide.

PCR Enzyme Selection Guide

Application

PCR Enzyme Properties

GC/AT-rich

complex template

TransTaq HiFi

TransStart seriesGC Enhancer

TransTaq HiFi

TransStart series

TransDirect series

routine PCR high fidelity PCR

TransTaq series

TransStart TaqTransPfu series

Easy Taq

TransFast Taq

high specificity PCR

TransStart series

long PCR

PCR inhibitor

enriched template

PCR Enzyme TransFastTaq EasyTaq TransTaq-T TransTaqHiFi TransStartTaq TransStartTopTaq

Amplication Efciency TransFast

Taq =EasyTaq

TransTaq-T TransTaqHiFi TransStartTaq TransStartTopTaq

Specicity TransFast

Taq =EasyTaq

TransTaq

-T TransTaq

HiFi TransStart

Taq TransStart

TopTaq

Fidelity(vs EasyTaq) 1 1 18 18 18 18

Extension Rate 6 kb/min 1-2 kb/min 1-2 kb/min 1-2 kb/min 1-2 kb/min 1-2 kb/min

Hot start - - + + + +

3-5 exo + + + + + +

Product Size (human

genomic DNA as

template

4 kb 4 kb 8 kb 12 kb 12 kb 12 kb

Specication


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High quality products

Applications

PCR Enzyme Applications

TransFastTaqDNA Polymerase routine PCR; fast PCR

EasyTaq DNA Polymerase routine PCR

TransTaq-T DNA Polymerase highly complex templates; TA cloning

TransTaqHiFi DNA Polymerase GC/AT; complex templates; long PCR; TA cloning

TransStartTaqDNA Polymerase qPCR; multiplex PCR; TA cloning

TransStartTopTaqDNA Polymerase qPCR; multiplex PCR; TA cloning

EasyPfuDNA Polymerase high delity PCR; blunt cloning; site-directed mutagenesis

TransStartFastPfuDNA Polymerase high delity PCR; fast PCR; robust PCR; blunt cloning; site-directed mutagenesis

TransStartFastPfuFlyDNA Polymerase high delity PCR; fast PCR; robust PCR; blunt cloning; site-directed mutagenesis

PCR Enzyme EasyPfu TransStart FastPfu TransStartFastPfu Fly

Amplication Efciency EasyPfu


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Description

TransFastTaq DNA polymerase is an engineered version ofTaq

DNA polymerase. The enzyme consists of a single polypeptide with a

molecular weight of approximately 94 kDa.

TransFastTaq DNA polymerase has 5-3 DNA polymerase activity

and 5-3 exonuclease activity. It lacks 3-5 exonuclease activity.

The extension rate is about 6 kb/min.

Template-independent A can be generated at the 3 end of the PCR

product. PCR products can be cloned in TA cloning vector.

Amplication of DNA fragment up to 4 kb.

Unit Denition

One unit ofTransFastTaqDNA Polymerase incorporates 10 nmol

of deoxyribonucleotide into acid-preceptible material in 30 minutes at

74oC.

Appl ications

Routine PCR.

Fast PCR.

Colony PCR.

Quality Contro l

TransFastTaqDNA polymerase has passed the following quality

control assays: functional absence of double-and single-stranded

endonuclease activity; >99% homogeneous measured by SDS-PAGE

gel electrophoresis. Each batch ofTransFastTaq DNA Polymerase

is assayed for amplication efciency from as little as 10 ng of human

genomic DNA.

Component Volume Final ConcentrationTemplate DNA Variable as required

Forward Primer (10 M) 1-2 l 0.2-0.4 M each

Reverse Primer (10 M) 1-2 l 0.2-0.4 M each

10TransFastTaq Buffer 5 l 1

2.5 mM dNTPs 4 l 0.2 mM

TransFastTaq DNA Polymerase 0.5 -1 l 2.5-5 units

ddH2O Variable -

Total volume 50 l -

Reaction Components

PROTOCOL

TransFastTaqDNA Polymerase

dNTPs

(2.5 mM)

AP101-11 500 units

AP101-12 6500 units

Concentration

5 units/l

Components

TransFastTaqDNA Polymerase

10TransFast Taq Buffer (200 mM Tris-HCl

pH 8.4; 100 mM KCl; 100 mM (NH4)2SO4;

20 mM MgSO4; others)

6DNA Loading Buffer

Storage

at -20oC for two years

dNTP-freeAP101-01 500 units

AP101-02 6500 units


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Description

EasyTaq DNA polymerase is purifed from E. coli expressing a cloned

Thermus aquaticus DNA polymerase. The enzyme consists of a

single polypeptide with a molecular weight of approximately 94 kDa.

EasyTaq

DNA polymerase has 5-3 DNA polymerase activity and 5-3

exonuclease activity. lt lacks 3-5 exonuclease activity. EasyTaq DNA

polymerase is suitable for routine amplication. PCR products are not

suitable for PAGE.

The extension rate is about 1-2 kb/min.



Amplication of DNA fragments up to 4 kb.

Appl ication

Routine PCR.

Unit Denition

One unit ofEasyTaqDNA Polymerase incorporates 10 nmol of into acid-

preceptible material in 30 minutes at 74oC.

EasyTaqDN A Polymerase

Concentration

5 units/l

Components

EasyTaq - DNA Polymerase

10EasyTaq Buffer (200mM Tris-HCl

pH8.3; 200 mM KCl; 100 mM (NH4)2SO4;


6DNA Loading Buffer

Storage

at -20oC for two year

Amplification Rate

0 50 100 150 200 250 300

1 Kb

2 Kb

3 Kb

4 Kb

min

EasyTaq

TransFast Taq

EasyTaq and TransFast Taq

Comparison Between

M ME F E F E F E F E F E F E F E F

M1Kb Plus DNA Ladder

EEasyTaq DNA Polymerase

FTransFast Taq DNA Polymerase

94oC 3 min

94oC 5 sec

50-60oC 15 sec 30-35 cycles

72o

C x sec72oC 5-10 min

0-2 kb

2-3 kb

>3 kb

10 sec/kb

20 sec/kb

30 sec/kb

dNTP-free

AP111-01 500 units

AP111-02

AP111-03

6500 units

42500 units

dNTPs

(2.5 mM)

AP111-11 500 units

AP111-12

AP111-13

6500 units

42500 units

Thermal cycling conditions

Target Extension time


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EasyTaqDNA Polymerase for PAGE

Concentration

5 units/l

94oC 2-5 min

94oC 30 sec


72oC 1-2 kb/min

72oC 5-10 min

M: 1Kb Plus DNA Ladder

1:CCRD 0.5 kb; 2: BDNF 0.8 kb;

3:Rhod 1.2 kb; 4: Rhod 2 kb;

5:Rhod 4.17 kb.

50 ng of Human Genomic DNA as templates

Component Volume Final Concentration

Template DNA Variable as required



10EasyTaq

Buffer 5 l 1


EasyTaq DNA Polymerase 0.5-1 l 2.5-5 units

ddH2O Variable -

Total volume 50 l -

Notes

MgSO4 is included in the 10PCR Buffer

at a nal concentration of 2.0 mM, which is

sufcient for most amplications. If needed,

suggested to use the 100 mM MgSO4

stock to prepare a titration from 2.0 mM to

4.0 mM (final concentration) in 0.25 mM

increments.

2.5 units ofEasyTaq DNA Polymerase is

enough for most target amplifications. For

some amplifications, up to 5 units/reaction

EasyTaq

DNA Polymerase can be used.

Reaction Components

PROTCOL


Quality Control

EasyTaq DNA polymerase has passed the following quality control

assays: functional absence of double- and single-stranded


gel electrophoresis. EasyTaq DNA Polymerase is assayed foramplication efciency from as little as 10 ng of human genomic DNA.

dNTP-free

(2.5 mM)

AP112-01 2500 units

AP112-02 42500 units

dNTPs

(2.5 mM)

AP112-11 2500 units

AP112-12 42500 units


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Components

EasyTaqDNA Polymerase for PAGE

10EasyTaq Buffer for PAGE (200mM

Tris-HCl pH8.3; 200 mM KCl; 100 mM

(NH4)2SO4; 20 mM MgSO4; others)

6DNA Loading Buffer

Storage


Description

EasyTaq DNA polymerase for PAGE is purifed from E. coli expressing

a cloned Thermus aquaticus DNA polymerase. The enzyme consists

of a single polypeptide with a molecular weight of approximately 94

kDa. EasyTaq

DNA polymerase for PAGE has 5-3 DNA polymeraseactivity and 5-3 exonuclease activity. lt lacks 3-5 exonuclease activity.

EasyTaq

DNA polymerase is suitable for routine amplication.





Appl ication

Short fragment PCR.

Unit Denition

One unit ofEasyTaq

DNA Polymerase for PAGE incorporates 10 nmolof deoxyribonucleotide into acid-preceptible material in 30 minutes at

74oC.

94oC 2-5 min

94oC 30 sec


72oC 1-2 kb/min

72oC 5-10 min





10EasyTaqBufferfor PAGE 5 l 1


EasyTaqDNA Polymerase for PAGE 0.5-1 l 2.5-5 units

ddH2O Variable -

Total volume 50 l -

Reaction Components

PROTOCOL


Quality Contro l

EasyTaq DNA polymerase for PAGE has passed the following quality

control assays: functional absence of double- and single-stranded


gel electrophoresis. EasyTaq DNA Polymerase for PAGE is assayed

for amplication efciency from as little as 10 ng of human genomic

DNA.


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Description

TransTaq-T DNA Polymerase is an engineered version ofTaq DNA

polymerase. TransTaq-T DNA Polymerase has end transferase activity

and exonuclease activity. TransTaq-T DNA Polymerase combines the

amplication efciency ofTaq DNA polymerase with the delity ofPfu

DNA polymerase.

dNTP-free

AP122-01 250 units

AP122-02 500 units

AP122-03 6500 units

dNTPs(2.5 mM)

AP122-11 250 units

AP122-12 500 units

AP122-13 6500 units


Concentration

5 units/l

Components


10TransTaq-TBuffer (200 mM

Unit Denition

One unit ofTransTaq-T DNA Polymerase incorporates 10 nmol of

deoxyribonucleotide into acid-preceptible material in 30 minutes at 74oC.

Quality Control

TransTaq

-T DNA polymerase has passed the following qualitycontrol assays: functional absence of double-and single-stranded


gel electrophoresis. Each batch ofTransTaq-T DNA Polymerase is

assayed for amplication efciency from as little as 10 ng of human

genomic DNA.


Template DNA Variable as requiredForward Primer (10 M) 1-2 l 0.2-0.4 M each


10TransTaq-TBuffer 5 l 1


TransTaq-T DNA Polymerase 0.5-1 l 2.5-5 units

ddH2O Variable -

Total volume 50 l -

Notes


at a nal concentration of 2.0 mM, which

is sufcient for most amplications. If

needed, suggested to use the 100 mM

MgSO4 stock to prepare a titration from 2.0

mM to 4.0 mM (nal concentration) in 0.25

mM increments.

2.5 units ofTransTaq-T DNA polymerase

is enough for most target amplications.

For some amplications, up to 5 units/

reaction TransTaq -T DNA polymerase

can be used.

Reaction Components

PROTOCOL

Fidelity is 18 times higher than EasyTaqDNA Polymerase.


product. PCR products can be cloned in TA cloning vector. The extension rate is about 1-2 kb/min.


Appl ications

Complex PCR.

TA cloning.

Tris-HCl pH9.0; 100 mM KCl; 100 mM

(NH4)2SO4; 20 mM MgSO4; others)

6DNA Loading Buffer

Storage



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Description

TransTaqDNA Polymerase High Fidelity (TransTaqHiFi DNA Polymerase)

isTransTaq

-T DNA Polymerase mixed with an 3-5exonuclease. TransTaq

HiFiDNA polymerase provides higher specicity and amplication efciency

thanTransTaq-T DNA Polymerase. The delity ofTransTaqHiFi DNA

polymerase is similar to EasyPfu DNA Polymerase.

Fidelity is 18 times higher than EasyTaq DNA Polymerase.



product. PCR products can be cloned in TA cloning vector


Appl ications

High delity.

High yield.

TransTaqHiFi Buffer I for genomic DNA, TransTaqHiFi Buffer II for

DNA, cDNA, Plasmid DNA amplication.

Unit Denition

One unit ofTransTaq HiFi DNA Polymerase incorporates 10 nmol of

deoxyribonucleotide into acid-preceptible material in 30 minutes at

74oC.

TransTaqDNA Polymerase High FidelityHiFi

Concentration

5 units/l

Components

TransTaqHiFi DNA Polymerase

10TransTaq HiFi Buffer I, II (200

mM Tris-HCl pH 9.0; 100 mM KCl; 100

mM (NH4)2SO4; 20 mM MgSO4; 10%

Glycerol,others)

6DNA Loading Buffer

GC Enhancer

Storage


dNTPs

(2.5 mM)

AP131-11 250 units

AP131-12 500 units

AP131-13 6500 units

dNTP-free

AP131-01 250 units

AP131-02 500 units

AP131-03 6500 units

94oC 2-5 min

94oC 30 sec


72

o

C 1-2 kb/min72oC 5-10 min

1 2 3 4 5 6 7M


1:BDNF 0.8 kb; 2: Rhod 1.2 kb;

3:-globin 1.3 kb; 4: Rhod 2.0 kb ;

5:-globin 3.0 kb; 6: Rhod 4.17 kb;

7: Factor IX 7.5 kb


M



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Notes MgSO4 is included in the 10PCR Buffer






mM increments.

2.5 units ofTransTaqHiFi DNA

polymerase is enough for most target

amplications. For some amplications,

up to 5 unit/reaction TransTaqHiFi DNA

polymerase can be used.

PROTOCOL

Reaction Components

Quality Control

TransTaqHiFi DNA polymerase has passed the following quality



gel electrophoresis. Each batch ofTransTaq

HiFi DNA Polymeraseis assayed for amplication efciency from as little as 10 ng of human

genomic DNA.

Components Volume Final Concentration

Template DNA variable as required



10TransTaqHiFi Buffer I/II 5 l 1


TransTaq

HiFi DNA Polymerase 0.5-1 l 2.5-5 units

ddH2O to 50 l Not applicable

M1: 1Kb Plus DNA Ladder

M2:Trans15K DNA Marker

Lane 1: REPA 1.8 kb; Lane 2: NCBP 2.5 kb;

Lane 3: HDP 3.5 kb; Lane 4: VIN 4.6 kb;

Lane 5: Pol 6.8 kb; Lane 6: APC 8.5 kb;

Lane 7: Dynein 12.3 kb

100 ng Human total RNA as templates

TransTaq

HiFi Buffer II

TransTaq

HiFi Buffer I

M1: 1Kb Plus DNA LadderM2:Trans15K DNA MarkerLane 1: BDNF 0.8 kb;

Lane 2: Rhod 1.2 kb

Lane 3: Rhod 2.0 kb;

Lane 4: -globin 3.0 kb

Lane 5: Rhod 4.17 kb;

Lane 6: Factor IX 7.5 kb

Lane 7: Serum albumin 12.4 kb

50 ng Human Genomic DNA as templates

Thermal cycling conditions94oC 2-5 min

94oC 30 sec


72oC 1-2 kb/min

72oC 5-10 min


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10TransTaqHiFi Buffer I/II 5 l 1


TransTaqHiFi DNA Polymerase 0.5-1 l 2.5-5 units

ddH2O Variable -

Total volume 50 l -

Notes MgSO4 is included in the 10PCR Buffer






mM increments.

2.5 units ofTransTaqHiFi DNA

polymerase is enough for most target

amplications. For some amplications,

up to 5 unit/reaction TransTaqHiFi DNA

polymerase can be used.

PROTOCOL

Reaction Components


94oC 30 sec


72oC 1-2 kb/min

72oC 5-10 min

TransStartTaqDNA Polymerase

Description

TransStartTaq DNA Polymerase is a hot start Taq DNA polymerase.

At room temperature, one binding protein binds to double-stranded

DNA template and another binding protein binds to primers. These

unique formulations effectively neutralize the DNA polymerase activity

at room temperature. The extension rate for TransStartTaq DNA

Polymerase is 1-2 kb/min. Template-independent A can be generated

at the 3 end of the PCR product. PCR products can be cloned in TA

cloning vector.

Appl ications Complex template.

GC/AT-rich template.

Multiplex PCR.

Unit Denition

One unit ofTransStartTaq DNA Polymerase incorporates 10 nmol of

deoxyribonucleotide into acid-insoluble material in 30 min at 74oC.

Quality Control

TransStartTaq DNA polymerase has passed the following quality



gel electrophoresis. Each batch of TransStartTaq DNA Polymerase


genomic DNA.

Concentration

2.5 units/l

Components

TransStartTaq DNA Polymerase

10TransStart TaqBuffer (500 mM Tris-

HCl pH 9.0; 200 mM (NH4)2SO4; 20 mM

MgSO4; 10% Glycerol; others)

6DNA Loading Buffer GC Enhancer

Storage


dNTP-free

AP141-01 250 units

AP141-02 500 units

AP141-03 6500 units

dNTPs

(2.5 mM)

AP141-11 250 unitsAP141-12 500 units

AP141-13 6500 units


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Notes


at a nal concentration of 2.0 mM, which is

sufcient for most amplications. If needed

suggested to use the 100 mM MgSO4

stock to prepare a titration from 2.0 mM to

4.0 mM (final concentration) in 0.25 mM

increments.

Choose GC Enhancer for the amplication

of GC/AT rich and complex template.

PROTOCOL

Reaction Components

TransStartHot Start ( Double Blocking )

TransStartdouble blocking technique blocks primer-dimer formation and polymerase activity at room temperature by blocking primers and DNA

templates at room temperature. Blocking proteins are released during the initial denaturation. This double blocking method has higher efciency

than antibody based, or chemical modied hot start PCR.





10TransStartTaq Buffer 5 l 1


TransStartTaq DNA Polymerase 0.5-1 l 1.25-2.5 units

ddH2O Variable -

Total volume 50 l -



1: REPA 1.8 kb 2: NCBP 2.5 kb

3: HDP 3.5 kb 4: VIN 4.6 kb

5: Pol 6.8 kb 6: APC 8.5 kb

7: Dynein 12.3 kb

Human cDNA as templates

TransTaq

HiFi Buffer IITransTaq HiFi Buffer I



1: BDNF 0.8 kb 2: Rhod 1.2 kb

3: Rhod 2.0 kb 4: -globin 3.0 kb

5: Rhod 4.17 kb 6: Factor IX 7.5 kb

7: Serum albumin 12.4 kb

50 ng of Human Genomic DNA as

templates


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94oC 30 sec


72oC 1-2 kb/min

72oC 10 minM1 1Kb Plus DNA LadderM2 Trans15K DNA Marker

1 Numb 0.3 kb 2 CCRD 0.5 kb 3 BDNF 0.8 kb 4 Rhod 1.2 kb5 Rhod 2.0 kb 6 -globin 3.0 kb 7 Rhod 4.17 kb 8 Factor IX 7.5 kb9 Serum albumin 12.4 kb50 ng of Human Genomic DNA as templates

TransStartTopTaqDNA Polymerase

Description

TransStart

TopTaq DNA Polymerase is an engineered version ofTaq

DNA Polymerase combined with TransStartTechnique. TransStartdouble

blocking technique prevents primer-dimer formation and polymerase

activity at room temperature by blocking the primers and DNA templates

at room temperature. Blocking proteins are released from primers and

templates during the initial denaturation. This double blocking method has

higher efciency than antibody based, or chemical modied hot start PCR.

Compare with the TransStart

Taq DNA Polymerase, the TransStart

TopTaq DNA Polymerase has higher amplication efciency and

sensitivity. Fidelity is 18 times higher than EasyTat

DNA Polymerase.

The specicity is higher than antibody based chemical or modied hot

start DNA polymerases.

Reduced nonspecic amplication and primer dimers.

Different from Taq antibody, no risk of contamination from mammal

DNA.

Different from chemical modication , heating step no longer needed.


Appl ications Complex PCR.

Multiplex PCR.

High delity and specicity PCR.

High yield.Unit Denition

One unit ofTransStartTopTaq DNA Polymerase incorporates 10 nmol

of deoxyribonucleotide into acid-insoluble material in 30 min at 74oC.

Quality Control

TransStart

TopTaq DNA polymerase has passed the following quality


endonuclease activity; >99 % homogeneous measured by SDS-PAGE

gel electrophoresis. Each lot ofTransStart

TopTaq DNA Polymerase


genomic DNA.

Concentration

2.5 units/l

Components

TransStartTopTaq DNA Polymerase

10TransStartTopTaqBuffer (500 mM

Tris-HCl (pH 9.0); 200 mM (NH4)2 SO4;


6DNA Loading Buffer

GC Enhancer

Storage


dNTP-free

AP151-01 250 units

AP151-02 500 units

AP151-03 6500 units

dNTPs

(2.5 mM)

AP151-11 250 units

AP151-12 500 units

AP151-13 6500 units


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Notes


at a final concentration of 2.0 mM, which issufficient for most targets amplificaiton. For

some targets, adjust the Mg2+ from 2.0 mM to

4.0 mM (nal concentration) to ensure a good

amplication.

Reaction Components


94oC 2-5 min

94oC 30 sec


72oC 1-2 kb/min

72oC 5-10 min

M: Trans2K

Plus II DNA Marker

Lane 1: Rhod 1.2 kb Lane 2: Rhod 2.0 kb Lane 3: -globin 3.0 kb

Lane 4: -globin 4.1 kb Lane 5: Rhod 4.17 kb Lane 6: -globin 6.1 kb

Lane 7: Factor IX 7.5 kb Lane 8: IGF2R 8.9 kb Lane 9: Serum albumin 12.4 kb

1 2 3 4 5 6 7 8 9M

M: Trans2K

Plus DNA Marker

Lane 1: DMD1 0.3 kb Lane 2: Numb 0.3 kb

Lane 3: P53 0.5 kb Lane 4: P1P2 1.2 kb

Lane A: A company hot start polymerase

Lane T: TransStartTopTaq DNA Polymerase

1 2 3 4

M A T A T A T A T

50 ng Human Genomic DNA as templates





10TransStartTopTaq Buffer 5 l 1


TransStartTopTaq DNA Polymerase 0.5-1 l 1.25-2.5 units

ddH2O Variable -

Total volume 50 l -

PROTOCOL


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EasyPfuDNA Polymerase

Description

EasyPfuDNA Polymerase is an engineered version ofpfu DNA Polymerase

with enhanced yield and higher delity. EasyPfu polymerase possesses

a proofreading 3'-5' exonuclease activity. Its delity is 18 times higher

than EasyTaq DNA Polymerase. EasyPfu DNA Polymerase is suitable

for blunt-end cloning and site-directed mutagenesis. Amplification of

fragment up to 6 kb.

Appl ications High delity.

Blunt-end cloning.

Site-directed mutagensis.

Unit Denition

One unit ofEasyPfuDNA P olymerase incorporates 10 nmol of

deoxyribonucleotide into acid-insoluble material in 30 min at 74oC.

Quality Control

EasyPfu polymerase has passed the following quality control assays:

functional absence of double- and single-stranded endonuclease

activity, >99% homogeneous measured by SDS-PAGE gel

electrophoresis. Each batch ofEasyPfu DNA Polymerase is assayed for

amplication efciency from as little as 10 ng of human genomic DNA.

dNTP-free

AP211-01 250 units

AP211-02 500 units

AP211-03 6500 units

dNTPs

(2.5 mM)

AP211-11 250 units

AP211-12 500 units

AP211-13 6500 units

Concentration

2.5 units/l

Components

EasyPfuDNA Polymerase

10EasyPfuBuffer(200 mM Tris-HCl

pH 8.8; 100 mM (NH4)2SO4; 100 mM

KCl; 20 mM MgSO4; others)

50 mM MgSO4

6DNA Loading Buffer

Storage


PROTOCOL

Reaction Components





10EasyPfu Buffer 5 l 1


EasyPfu DNA Polymerase 1 l 2.5 units

ddH2O Variable -

Total volume 50 l -


94oC 2-5 min

94oC 30 sec


72oC 0.5 kb/min

72oC 5-10 min


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1:CCRD 0.5 kb; 2: BDNF 0.8 kb; 3: Rhod 1.2 kb; 4: -globin 1.3 kb;

5: Rhod 2 kb; 6: -globin 3 kb; 7: Rhod 4.17 kb; 8: -globin 4.1 kb


1 2 3 4 5 6 7 8M M

TransStart

FastPfuDNA Polymerase

Description

TransStartFastPfu DNA Polymerase is a hot start, high fidelity,

and high processivity DNA polymerase. The fidility of TransStart

FastPfu DNA polymerase is 54-fold higher than TaqDNA polymerase.TransStart

FastPfu DNA polymerase has extension rate up to 4 kb/

min. PCR product amplied by TransStartFastPfu DNA polymerase is

blunt end and can be cloned into blunt cloning vector.

Appl ications

High delity PCR.

High yield and robust PCR.

Blunt end cloning.

Site-directed mutagenesis.

Unit Denition

One unit of TransStart FastPfu DNA Polymerase incorporates 10

nmol of deoxyribonucleotide into acid-insoluble material in 30 min

at 74oC.

Quality Control

TransStartFastPfu DNA Polymerase has passed the following quality



gel electrophoresis. Each lot ofTransStartFastPfu DNA Polymerase


genomic DNA.

dNTP-free

AP221-01 250 units

AP221-02 500 units

AP221-03 6500 units

dNTPs

(2.5 mM)

AP221-11 250 units

AP221-12 500 units

AP221-13 6500 units

Concentration

2.5 units/l

Components

TransStartFastPfu DNA Polymerase

5TransStartFastPfu Buffer (100 mM

Tris-SO4 pH 9.2; 50 mM (NH4)2SO4; 200

mM KCl; 10 mM MgSO4; 10% Glycerol;

others)

50 mM MgSO4

6DNA Loading Buffer

PCR Stimulant

Storage



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Suggested conditions

50 lreaction volume

Parameter Targets10 kb Targets10 kb cDNA Targets

Template100 ng Genomic DNA 200-500 ng Genomic DNA 1-2 l cDNA from RT reaction

5-30 ng Plasmid DNA 5-30 ng Plasmid DNA (50-500 ng starting RNA template)

MgSO4 Add 1-2 l of 50 mM MgSO4 to a nal concentration of 3-4 mM for target larger than 5 kb


Number of cycles Temperature Plasmid or Genomic DNA cDNA

1 95oC 2 min 1 min

Plasmid or Genomic DNA: 30-35

cDNA: 40

95oC 20 sec 20 sec

Tm-5oC 20 sec 20 sec

72oC15 sec for targets1 kb

15-30 sec per kb for targets >1 kb

30 sec for targets1 kb

30 sec per kb for targets>1 kb

1 72oC 5 min 5 min

Reaction Components





5TransStartFastPfu Buffer 10 l 1


TransStartFastPfu DNA Polymerase 1 l 2.5 units

ddH2O Variable -

Total volume 50 l -

cDNA amplication with TransStartFastPfu DNA Polymerase

1 2 3 4 5M1 M2

M1 1Kb Plus DNA Ladder

M2 Trans15K DNA Marker

1 ACTR 3.5 kb;

2 VIN 4.6 kb;

3 Pol 6.8 kb;

4 APC 8.5 kb;

5 Dynein 12.3 kb;

2 hrs 14 min

2 hrs 34 min

3 hrs 09 min

3 hrs 41 min

4 hrs 48 min


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M: Trans15K DNA Marker

1:UDG 7.0 kb;

2:LN 10.0 kb;

3:Fang 14.7 kb;

Genomic DNA amplication with TransStartFastPfu DNA Polymerase

1 2 3 4 5M1 M2



1 Rhod 2.0 kb;

2 -globin 3.0 kb;

3 Rhod 4.17 kb;

4 Factor IX 7.5 kb;

5 Serum albumin 12.4 kb;

1 hrs 19 min

1 hrs 27 min

1 hrs 29 min

3 hrs 25 min

4 hrs 48 min

1 2 3M M

Plasmid DNAamplication with TransStartFastPfu DNA Polymerase

1 hrs 36 min

1 hrs 55 min

2 hrs 26 min

TransStartFastPfuFly DNA Polymerase

dNTP-free

AP231-01 250 units

AP231-02 500 units

AP231-03 6500 units

dNTPs

(2.5 mM)

AP231-11 250 unitsAP231-12 500 units

AP231-13 6500 units

Concentration

2.5 units/l

Components

TransStartFastPfu Fly DNA Polymerase

5TransStartFastPfu FlyBuffer (100

mM Tris-SO4 pH 9.2; 50 mM (NH4)2SO4;

200 mM KCl; 10 mM MgSO4; 10%

Glycerol; others)

Description

TransStartFastPfu Fly DNA Polymerase is a hot start, high delity, and

high processivity DNA polymerase. The dility of TransStartFastPfu

Fly DNA polymerase is 108-fold higher than TaqDNA polymerase.

TransStartFastPfu Fly DNA polymerase has extension rate up to 6 kb/

min. PCR product amplied by TransStartFastPfu FlyDNA polymerase

is blunt end and can be cloned into blunt cloning vector.

Appl ications

High delity PCR.

GC rich template and template that has secondary struture.

High yield and robust PCR.

Blunt end cloning.


Long-segment PCR.

50 mM MgSO4

6DNA Loading Buffer

PCR Stimulant

Storage



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Unit Denition

One unit of TransStart FastPfu Fly DNA Polymerase incorporates

10 nmol of deoxyribonucleotide into acid-insoluble material in 30

min at 74oC.

Quality Contro lTransStartFastPfu Fly DNA Polymerase has passed the following

quality control assays: functional absence of double- and single-

stranded endonuclease activity; >99% homogeneous measured by

SDS-PAGE gel electrophoresis. Each lot ofTransStartFastPfu DNA

Polymerase is assayed for amplication efciency from as little as 10

ng of human genomic DNA.

Suggested conditions

50 lreaction volume

Parameter Targets10 kb Targets10 kb cDNA Targets

Template100 ng Genomic DNA 200-500 ng Genomic DNA 1-2 l cDNA from RT reaction

5-30 ng Plasmid DNA 5-30 ng Plasmid DNA (50-500 ng starting RNA template)

MgSO4 Add 1-2 l of 50 mM MgSO4 to a nal concentration of 3-4 mM for target larger than 5 kb


Number of cycles Temperature Plasmid or Genomic DNA cDNA

1 95oC 2 min 1 min

Plasmid or Genomic DNA: 30-35cDNA: 40

95oC 20 sec 20 sec

Tm-5

o

C 20 sec 20 sec

72oC15 sec for targets1 kb

15-30 sec per kb for targets >1 kb

30 sec for targets1 kb

30 sec per kb for targets>1 kb

1 72oC 5 min 5 min

Reaction Components





5TransStartFastPfu FlyBuffer 10 l 1


TransStartFastPfu Fly DNA Polymerase 1 l 2.5 units

ddH2O Variable -

Total volume 50 l -

1

18

54

108

1X

18X

108X

54X

EasyPfu TranStart

FastPfu

TranStart

FastPfuFlyEasyTaq

Fide

lity


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Description

GC Enhancer is used with DNA Polymerase, to optimize PCR of complex

templates or GC/AT-rich templates. The stock concentration is 10,

and the working concentration can be varied between 0.5- 5.

Appl ication

Amplication of complex templates and/or GC/AT-rich templates.

GC Enhancer

(50 l reaction mixture as example)

Storage


AG101-01 200 l

GC Enhancers Volume (l) Final Concentration

2.5 0.5

5 1

10 2

15 3

20 4

25 5

10 kb

7 kb

4 kb

Size

PCR Time4 kb: Genomic DNA; 7 kb and 10 kb: Plasmid DNA

Total Reaction Time


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PCR Stimulant

DescriptionPCR Stimulant is used to optimize PCR of complex templates or GC/

AT-rich templates. It is especially suitable forPfu enzymes. The stock

concentration is 5, and the working concentration can be varied

between0.5- 2.5.

Appl ication

Amplication for complex templates and/or GC/AT-rich templates.

Storageat -20oC for two years

AG111-01 200 l

(50 l reaction volume as example)

PCR Stimulants Volume (l) Final Concentration

5 0.5

10 1

20 2

25 2.5


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PCR Enzyme vs PCR SuperMix

PCR SuperMix(+dye)

electrophoresis

PCR SuperMix(-dye)

cloning and sequencing

PCR SuperMix Selection Chart

optimize the PCR system

simple and quick

PCR Enzyme PCR SuperMix

purify

PCR SuperMixesPCR SuperMix is a ready-to-use mixture of DNA polymerase, salt, magnesium, dNTPs, and other components for efcient

PCR amplication. All you have to do is to add template, primers, and dH2O. If using PCR SuperMixes with dye, dye needs

to be removed before sequencing.

Appl ications

PCR SuperMix Application

2EasyTaqPCR SuperMix routine PCR

2TransTaq-T PCR SuperMix complex templates and TA cloning

2TransTaqHigh Fidelity(HiFi)

PCR SuperMix I

GC/AT-rich templates, complex templates

long fragment amplication, genomic DNA amplication (


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PROTOCOL

Total volume 50 l -

EasyTaq

PCR SuperMix (+dye) EasyTaq

PCR SuperMix (-dye)

M


1:CCRD 0.5 kb

2:BDNF 0.8 kb

3:Rhod 1.2 kb

4: Rhod 2 kb

5:Rhod 4.17 kb

50 ng of Human Genomic DNA as

templates

M M 1 2 3 4 51 2 3 4 5





2EasyTaqPCR SuperMix 25 l 1

ddH2O Variable -

Notes

Completely thaw the contents in the tube

before each use.

Reaction Components


94oC 2-5 min

94oC 30 sec


72oC 1-2 kb/min

72oC 5-10 min

Description

EasyTaq PCR SuperMix is a ready-to-use mixture ofEasyTaq

DNA Polymerase, dNTPs, and optimized buffer. This SuperMix

is provided at 2 concentration and used at 1 concentration

by adding template, primers and H2O. PCR products are not

suitable for PAGE.

The extension rate is about 1-2 kb/min. Template-independent A can be generated at the 3 end of the PCR



Appl ication

Routine PCR.

Mix(-dye)

AS111-01 1 ml

AS111-02 51 ml

AS111-03 151 ml

Mix(+dye)

AS111-11 1 ml

AS111-12 51 ml

AS111-13 151 ml

2EasyTaqPCR SuperMix

Storage



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PROTOCOL


Template DNA

Variableas required



2EasyTaqPCR SuperMix for PAGE 25 l 1

ddH2O Variable -

Total volume 50 l -

Notes


before each use.

Reaction Components


94oC 2-5 min

94oC 30 sec


72oC 1-2 kb/min

72oC 5-10 min

Description

EasyTaq PCR SuperMix for PAGE is a ready-to-use mixture of

EasyTaq DNA Polymerase for PAGE, dNTP s, and optimized

buffer.This SuperMix for PAGE is provided at 2 concentration

and used at 1 concentration by adding template, primers and

H2O.





Appl ication

PCR of short fragments.

2EasyTaqPCR SuperMix for PAGE

Storage


Mix (+dye)

AS112-11 1 ml

AS112-12 51 ml

AS112-13 151 ml


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PCR SuperMix


Description

TransTaq-T PCR SuperMix is a ready-to-use mixture ofTransTaq-T

DNA Polymerase, dNTPs, and optimized buffer. This SuperMix is

provided at 2concentration and used at 1concentration by adding

template, primers and H2O.

Fidelity is 18 times higher than EasyTaq DNA Polymerase.





Appl ications

Complex template.

TA cloning.

Mix (-dye)AS122-01 1 ml

AS122-02 51 ml

Mix (+dye)AS122-11 1 ml

AS122-12 51 ml

2TransTaq-T PCR SuperMix

Storage

at -20OC for two years





2TransTaq-T PCR SuperMix 25 l 1

ddH2O Variable -


94 2-5 min

94 30 sec

50-60 30 sec 30-35 cycles

72 1-2 kb/min

72 5-10 min

PROTOCOL

Reaction ComponentsNotes


before each use.

TransTaq-T DNA Polymerase TransTaq

-T PCR SuperMix

MMM 1 2 3 4 5 6 7M: 1Kb Plus DNA Ladder

1:BDNF 0.8 kb; 2: Rhod 1.2 kb;

3:-globin 1.3 kb; 4: Rhod 2.0 kb;

5: -globin 3.0 kb; 6: Rhod 4.17 kb;

7: Factor IX 7.5 kb


1 2 3 4 5 6 7

Total volume 50 l -


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2TransTaqHiFi PCR SuperMix 25 l 1

ddH2O Variable -

Reaction Components

PROTOCOL

DescriptionTransTaq

High Fidelity (HiFi) PCR SuperMix I or II is a ready-to-

use mixture ofTransTaq High Fidelity(HiFi) DNA polymerase (a

hot start DNA polymerase ), with a proofreading 3-5 exonuclease,

dNTPs, and optimized buffer. TransTaq High Fidelity (HiFi) PCR

SuperMix I is designed for the amplification of genomic DNA, and

TransTaqHigh Fidelity (HiFi) PCR SuperMix II is designed for the

amplication of DNA, cDNA, or plasmid DNA. The delity is 18 fold

higher than EasyTaq DNA polymerase. This SuperMix is provided at 2

concentration and can be used at 1concentration by adding template,

primers and H2O.

Fidelity is 18 times higher than EasyTaq DNA Polymerase. The extension rate is about 1-2 kb/min.




Appl ications

High delity.

High yield.

Mix I(-dye)AS131-01 1 ml

AS131-02 51 ml

Mix II(-dye)AS131-21 1 ml

AS131-22 51 ml

2TransTaqHigh FidelityHiFiPCR SuperMix

Storage

at -20oC fortwo years

Notes

Completely thaw the contents in the tubebefore each use.


94oC 2-5 min

94oC 30 sec


72oC 1-2 kb/min

72oC 5-10 min

Total volume 50 l -


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PCR SuperMix


TransTaq

HiFi DNA Polymerase TransTaq

HiFi PCR S uperMix II



1:REPA 1.8 kb; 2: NCBP 2.5 kb; 3: HDP 3.5 kb; 4: VIN 4.6 kb;

5:Pol 6.8 kb; 6:APC 8.5 kb; 7: Dynein 12.3 kb


M21 2 3 4 5 6 7 M2M1 M1 1 2 3 4 5 6 7

TransTaq

HiFi DNA Polymerase TransTaq

HiFi PCR SuperMix I



1:BDNF 0.8 kb; 2:Rhod 1.2 kb; 3: Rhod 2.0 kb; 4: -globin 3.0 kb;

5:Rhod 4.17 kb; 6: Factor IX 7.5 kb; 7: Serum albumin 12.4 kb


M21 2 3 4 5 6 7 M2M1 M1 1 2 3 4 5 6 7


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Description

EasyPfuPCR SuperMix is a ready-to-use mixture ofEasyPfu DNA

polymerase , dNTPs, and optimized buffer. This SuperMix is provided

at 2concentration and used at 1concentration by adding template,

primers and H2O. PCR products can be cloned into pEASY-Blunt

cloning vector series.

Appl ications

High delity PCR amplication.

Blunt end cloning.



AS211-02 51 ml

2EasyPfuPCR SuperMix

Storage


PROTOCOL


94oC 2-5 min

94oC 30 sec


72oC 0.5 kb/min

72oC 5-10 min


1:CCRD 0.5 kb; 2: BDNF 0.8 kb; 3: Rhod 1.2 kb; 4: -globin 1.3 kb;

5: Rhod 2 kb; 6: -globin 3 kb; 7: Rhod 4.17 kb; 8: -globin 4.1 kb


1 2 3 4 5 6 7 8 M1 2 3 4 5 6 7 8

EasyPfu DNA Polymerase 2xEasyPfu PCR SuperMix

MM

Notes


before each use.Component Volume Final Concentration

Template DNA Variable


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PCR SuperMix


Description

TransStart

FastPfu PCR SuperMix is a ready-to-use mixture of

TransStartFastPfu DNA polymerase, dNTPs, and optimized buffer.

This SuperMix is provided at 2concentration and can be used at

1concentration by adding template, primers and H2O. PCR product

amplied by TransStartFastPfu DNA polymerase can be cloned into

pEASY-Blunt cloning vector series.

Extension rate up to 4 kb/min.

High delity, robust, and high speed amplication.

Appl ications

High delity amplication.


Long or complex template amplication.


AS221-02 51 ml

2TransStartFastPfu PCR SuperMix

Storage


PROTOCOL

Reaction Components


94oC 20 sec


72oC 2-4 kb/min

72oC 5-10 min

Notes


before each use.


Template DNA Variable


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Description

TransDirectTM Animal Tissue PCR Kit uses a propriety formulated

lysis reagent to release nucleic acids from a variety of animal

tissues(fresh,frozen). Unpurified DNA is used as template for PCR

using 2TransDirectTM PCR SuperMix which has extremely high

resistance to many PCR inhibitors found in animal tissues.

TransDirect

TM

Animal Tissue PCR Kit

Storage


Kit Components

Amount of Starting Mater ial

Component AD201-01 AD201-02

AD1 Buffer 12 ml 55 ml

AD2 Preparation Buffer 3 ml 15 ml

AD3 Buffer 12 ml 512 ml

2TransDirectTM PCR SuperMix(+dye) 1 ml 51 ml

ddH2O 5 ml 25 ml

Material Amount

Mammalian Cell 1-5106 cell

Animal Tissues 10-30 mg

Mouse Tail 0.5-1 cm sections

Mouse Ear 0.5-0.7 cm disk

Saliva 10-30 l

Hair shaft 30 mg

Appl ications

Propriety formulated lysis reagent to release nucleic acids from avariety of animal tissues like mammalian cells, saliva, hair shaft,

animal .

Extremely high animal tissue inhibitor resistance 2TransDirectTM

PCR SuperMix for direct PCR from unpuried DNA.

Amplications up to 3 kb.

AD201-01 100 rxns20 l system


Direct PCR Application

TransDirectTM Animal Tissue PCR Kit mammalian cells, saliva, hair shaft, animal tissues

TransDirectTM Plant Tissue PCR Kit Non polysaccharides and polyphenols in plant tissue

TransDirectTM Blood PCR KitFresh and frozen blood sample stored in EDTA, heparin, or citric acid

Blood sample and dried blood without the anticoagulant

Direct PCRTransDirect PCR uses a propriety formulated lysis reagent to release nucleic acids from a variety of animal

tissues(fresh,frozen), plant tissues. Unpuried DNA is used as template for PCR using 2TransDirectTM PCR SuperMix which

has extremely high resistance to many PCR inhibitors found in animal tissues, plant tissues and blood.


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ProcedureNotes


before each use.


Tissue Extract 4 l as required

2TransDirectTM PCR SuperMix (+dye) 10 l 1

Forward Primer (10 M) 0.4-0.8 l 0.2-0.4 M each

Reverse Primer (10 M) 0.4-0.8 l 0.2-0.4 M each


A:Add 25 l of AD2 buffer into 100 l of the AD1 Buffer, mix thoroughlyby pipetting up and down (where substantial extracted sample, AD1

buffer to AD2 Buffer at ratio of 4:1, pre-mixed, and use within two

hours).

B:For different samples, the following processing methods:

Mammalian Cells

From collected cells, thoroughly remove the culture broth, add the

mixture of AD1 and AD2, mix thoroughly by pipetting up and down.

Animal Tissues

Cut using clean, sterile scissors or blade, add the mixture of AD1 and

AD2, mix thoroughly by pipetting up and down.

Saliva

Directly add saliva into the mixture of AD1 and AD2, mix thoroughly

by pipetting up and down.

Hair Shafts

Cut hair into pieces, add the mixture of AD1 and AD2, mix thoroughly

by pipetting up and down.

C:Incubate for 10 minutes at room temperature (for difficult to lyse

cells such as hair, 55oC incubated for 10 minutes, followed by 95oC

incubated for 10 minutes). After incubation, 3 minutes at 95 oC.

D: Adding 100 ul AD3 Buffer, mix directly as template for PCR, or placed

and stored at 4oC or -20oC.

Thermal cycling conditions94 5-10 min

94 30 sec

50-60 30 sec 35-40 cycles

72 1-2 kb/min

72 5-10 min

M

M:Trans2K

Plus II DNA Marker

Lane 1: Hair 0.8 kb

Lane 2: Saliva 0.8 kb

Lane 3: HeLa 0.8 kb

Lane 4: HeLa 2 kb

Lane 5: HeLa 3 kb

Lane 6: Mouse ear 0.86 kb

Lane 7: Mouse ear 1.8 kb

7 8 9 10 11 121 2 3 4 5 6 13 14

Lane 8: Mouse ear3 kb

Lane 9: Drosophila 0.42 kb

Lane 10: Drosophila 2 kb

Lane 11: Nematode 0.9 kb

Lane 12: Shrimp 1.1 kb

Lane 13: Crab 1 kb

Lane 14: Razor clan 0.56 kb


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Description

TransDirectTM Plant Tissue PCR Kit uses a propriety formulated

lysis reagent to release nucleic acids from a variety of plant tissues.

Unpurified DNA is used as template for PCR using 2TransDirectTM

PCR SuperMix which has extremely high resistance to many PCR

inhibitors found in plant tissues.

Appl ications

Propriety formulated lysis reagent to release nucleic acids from a

variety of plant tissues

Extremely high plant tissue inhibitor resistance 2TransDirectTM

PCR SuperMix for direct PCR from unpuried DNA or PCR from cell

culture without DNA extraction.Amplications up to 2 kb.

TransDirectTM Plant Tissue PCR Kit

Storage


PROTOCOL

Notes


before each use.



Components AD301-01 AD301-02

PD1 Buffer 12 ml 55 ml

PD2 Buffer 12 ml 55 ml

2TransDirectTM PCR

SuperMix (+dye)1 ml 51 ml

ddH2O 5 ml 25 ml

Kit Components

Incubate the bottle for PD1 at 55oC until it becomes clear, if it is

viscous

A: Genomic DNA Extraction

1.Cut up 10-30 mg or 1cm2 plant tissue and add it to a tube containing

100 l of PD1 buffer, Vortex.

2.Incubate at 95C for 10 min.

3.Add 100 l of PD2 Buffer and vortex to mix. DNA is ready for PCR

or storage (Tissue Extract can be stored at 4oC for six months, or at

-20oC one year) .

B:PCR amplication


Tissue Extract 2-4 l as required

2TransDirectTM PCR SuperMix (+dye) 10 l 1





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Description

Trans Blood PCR SuperMix is a ready-to-use mixture ofTransBD

Taq DNA Polymerase, dNTPs, and optimized buffer. This SuperMix is

provided at 2concentration and can be used at 1concentration by

adding blood, primers and H2O.

Appl ications

Fresh and frozen blood sample stored in EDTA, heparin, or

citric acid.

Blood sample and dried blood without the anticoagulant.

PCR from cell culture without DNA extraction.

Amplication upto 4 kb.

Mix (-dye)AD401-01 100 rxns20 l system


TransDirectTM Blood PCR Kit

Storage


Thermal cycling conditions94 2-5 min

94 20 sec

50-60 20 sec 30-35 cycles

72 2-4 kb/min

72 5-10 min

M 7 8 9 10 11 121 2 3 4 5 6 13 14 15 16 17 18 19 20

M:Trans2K

Plus II DNA Marker

Lane 1: Corn 0.4 kb

Lane 2: Corn 0.8 kb

Lane 3: Corn 1.5 kb

Lane 4: Wheat 0.4 kb



Lane 7: Soybean 0.9 kb

Lane 8: Soybean1.5 kb

Lane 9: Arabidopsis 0.5 kb



Lane 12: Tobacco 0.5 kb

Lane 13: Tobacco 0.9 kb

Lane 14: Tobacoo 1.5 kb

Lane 15: Cotton 0.3 kb

Lane 16: Cotton 1 kb

Lane 17: Cotton 1.6 kb

Lane 18: Rice 0.3 kb

Lane 19: Rice0.9 kb

Lane 20: Rice 1.9 kb

Kit Components

Components AD401-01 AD401-02

2TransDirectTM PCR

SuperMix (+dye)1 ml 51 ml

ddH2O 5 ml 25 ml


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human frozen EDTA anticoagulated blood

human frozen heparin anticoagulated blood

human fresh blood sample without the anticoagulant

0.5 l of blood sample was used in 25 l PCR reaction.

Hela cells were used to amplify the Hdtgene (0.32 kb) in 25 l PCR reaction

M: Trans2K

DNA Marker

Lane 1: 104cells

Lane 2: 103cells

Lane 3: 102cells

Lane 4: 10 cells

Human oral cavity epithelial tissue wasused to amplify the Hdt gene (0.32 kb)

in 25 l reaction

M: Trans2K

DNA Marker

0.5 l of 1:40 diluted mouse heparin

anticoagulated blood was used in

25 l reaction .

M: Trans2K

DNA MarkerLane 1: Neo 0.25 kb

Lane 2: MAP 0.6 kb

0.5 l of diluted chicken sodium citrate

anticoagulated blood was used to amplifya 0.25 kb fragment in 25 l reaction

M: Trans2K

DNA MarkerLane 1: Blood

Lane 2: 1:10 dilution

Lane 3: 1:100 dilution

PROTOCOL

Reaction Components


94oC 5 min

94oC 30 sec


72o

C 1-2 kb/min72oC 5-10 min

Notes


before each use.


Template 0.2-0.8 l as required



2TransDirectTMPCR SuperMix (+dye) 10 l 1



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RT-PCR

Transreverse transcriptase series include

EasyScript RT,TransScript RT andTransScript II RT.

Products Size ofproduction Sensitivity FidelityComplex or GC-rich

template

EasyScriptRT 8 kb

TransScriptRT 12 kb

TransScript

II RT 15 kb

EasyScriptFirst-Strand cDNA Synthesis SuperMix 8 kb

TransScriptFirst-Strand cDNA Synthesis SuperMix 12 kb

TransScriptOne-Step gDNA Removal and cDNA

Synthesis SuperMix12 kb

TransScriptAll-in-One First-Strand cDNA Synthesis

SuperMix for PCR12 kb


SuperMix for qPCR12 kb

TransScriptII First-Strand cDNA Synthesis SuperMix 15 kb

TransScript II One-Step gDNA Removal and cDNA

Synthesis SuperMix 15 kb

TransScript II All-in-One First-Strand cDNA Synthesis

SuperMix for PCR15 kb

TransScript II All-in-One First-Strand cDNA Synthesis

SuperMix for qPCR15 kb

TransScriptTwo-Step RT-PCR SuperMix 12 kb

TransScript II Two-Step RT-PCR SuperMix 15 kb

EasyScriptOne-Step RT-PCR SuperMix 4 kb

TransScriptOne-Step RT-PCR SuperMix 8 kb

TransScript II One-Step RT-PCR SuperMix 8 kb

TransScript

II RT

TransScript RT

EasyScript

RT

Trans reverse transcriptases

Site mutation tohave a higherthermostability

Site mutation conveyinghigher synthesis ability

Site mutation todestroy RNase Hactivity


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U A

NNNNGAAAAAAAAAAAAAAA+GTTTTTTTTTTTTC C

NNNNUAAAAAAAAAAAAAAAAATTTTTTTTTTTT

NNNNCAAAAAAAAAAAAAAAAGTTTTTTTTTTTT

NNNNGAAAAAAAAAAAAAAAA

CTTTTTTTTTTTT

UNNNNCAAAAAAAAAAAAAAAA+TTTTTTTTTTTT

G

UNNNNCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

G TTTTTTTTTTTT

Trans RT products (except one-step kits) use Anchored Oligo(dT) to increase cDNA yield and full length cDNA

products

Anchored oligo(dT) binds specic site.

Higher cDNA yield and more full length cDNA.

Poly(A) tail can be a few hundreds bp long and oligo(dT)

binds randomly.

Difcult to synthesize long poly(A) tail.

Lower yield and less full length cDNA.

Anchored Oligo(dT) PrimerTraditionalOligo(dT)12-18 Primer

Primer

RNA

2RT Reaction Mix

RT/RI Enzyme Mix

Tradtional RT vs Trans RT

Mix RNA and primers

Denature 5 min at 65oC,

place on ice

Add buffer RNase

Inhibitor dNTPs

M-MLV RT

Incubate at 42oC for

50 min , then 15 min at

70oC to inactive the

enzyme

Add RNA primers

buffer RNase Inhibitor

dNTPs Trans RT,

in one tube

Incubate at 42oC/50oC

for 30 min , then 5 minat 85oC to inactive the

enzyme

Add all components

without thermal

denaturation and

ice-bath

Traditional First-Strand cDNA Synthesis vs TransFirst

-Strand cDNA Synthesis SuperMix

TradtionalTraditional TransTrans

Primer

RNA

Buffer

dNTPs

RT Enzyme

RNase Inhibitor


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EasyScript Reverse Transcriptase[M-MLV,RNase H ]

AE101-02 10000 units

AE101-03 510000 units

Description

EasyScript Reverse Transcriptase is an engineered version of

M-MLV reverse transcriptase with reduced RNase H activity. The

enzyme is purified to near homogeneity from E.coli containing the

modied M-MLV RT gene.

Reduced RNase H activity to reduce RNA template degradation

during the rst-strand cDNA synthesis.

Anchored oligo(dT)18 for higher yield and more full length cDNA.

cDNA up to 8 kb.

Appl ication

First strand cDNA synthesis.

Multiple copy gene.

Unit Denition

One unit ofEasyScript is the amount of enzyme required to

incorporate 1 nmol of deoxyribonucleotide into acid precipitable

material in 10 minutes at 37oC using Poly(A)Oligo(dT) as template

primer.

Concentration

200 units/l

Components

EasyScriptRT

5ES RT Buffer (375 mM KCl;

15 mM MgCl2; 100 mM Tris-HCl pH 8.4)

Anchored Oligo(dT)18

Storage

at -20oC for one year

PROTOCOL

First Strand cDNA synthesis1 Reaction Components

Component Volume

Total RNA/mRNA Variable

Anchored Oligo(dT)18(0.5 g /l) 1 l

or Random Primer(N9) (0.1 g/l) 1 l

or GSP 2 pmol

10 mM dNTPs 1 l

5ES RT Buffer 4 l

Ribonuclease Inhibitor (50 units/l) 0.5 l

EasyScript

RT 1 l

RNase-free Water to 20 l

2 Incubation

For anchored oligo(dT)18primer or GSP, incubate at 42oC for 30 min.

For random primer, incubate at 25oC for 10 min, thenat 42oC for 30 min.

3 Incubate at 85oC for 5 min to inactive enzyme.


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94oC 2-5 min

94

o

C 30 sec50-60oC 30 sec 35-40 cycles

72oC 1-2 kb/min

72oC 5-10 min

M 1Kb Plus DNA Ladder

1 GAPDH 0.5 kb 2 GAPDH 0.9 kb

3 REPA 1.8 kb 4 ACTR 3 kb

5 ACTR 3.5 kb 6 VIN 4.6 kb

7 TSC 5.3 kb 8 Pol 6.8 kb

Human cDNAas template

1 2 3 4 5 6 7 8M

NCBP Human DNA Polymerase

M M M1 2 3 4 5 6 7 1 2 3 4 5 6 7M 1Kb Plus DNA Ladder

Lanes 1-7 0 0.01 0.1 1 10 100 1000 pg

Human total RNA

M


cDNA Variable as requiredForward Primer (10 M) 1 l 0.2 M

Reverse Primer (10 M) 1 l 0.2 M

10TransTaqHiFi Buffer II (Mg2+

) 5 l 1


TransTaqHiFi DNA Polymerase 0.5 l 2.5 units

ddH2O Variable -

Total volume 50 l -

RT-PCRReaction Components


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AT101-02 10000 units

AT101-03 510000 units

TransScriptReverse Transcriptase[M-MLV,RNase H ]

Concentration

200 units/l

Components

TransScript RT

5TS RT Buffer (250 mM KCl;

15 mM MgCl2; 100 mM Tris-HCl pH 8.4)


Storage


NCBP

M M M

Human DNA Polymerase

1 2 3 4 5 6 7 1 2 3 4 5 6 7

M Trans15K DNA Marker

1 GAPDH 0.9 kb 2 REPA 1.8 kb

3 ACTR 3 kb 4 HDP 3.5 kb

5: VIN 4.6 kb 6 TSC 5.3 kb

7 Pol 6.8 kb 8 FIB 9.4 kb9 Dynein 12.3 kb

Human cDNA as template

M 1Kb Plus DNA Ladder

Lanes 1-7 0 0.01 0.1 1 10 100 1000 pg

Human total RNA

PROTOCOL

The condition for the rst strand cDNA synthesis is the same as for

EasyScript Reverse Transcriptase ( P39 ).

M1 2 3 4 5 6 7 8 9M

Description

TransScript Reverse Transcriptase is a recombinant M-MLV reverse

transcriptase with reduced RNase H activity.




cDNA up to 12 kb.

Appl ication


Multiple copy and low-copy genes.

Unit Denition

One unit ofTransScript is the amount of enzyme required to

incorporate 1 nmol of deoxyribonucleotide into acid-precipitable


primer.


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First Strand cDNA synthesis

1 Reaction Components

2 Incubation


For random primer, incubate at 25oC for 5-10 min then at 50oC for 30

min.

For GC-rich or complex secondary structure RNA template, incubate

at 55oC for 30 min.


PROTOCOL

Component Volume




or GSP 2 pmol

10 mM dNTPs 1 l

10TS II RT Buffer 2 l

Ribonuclease Inhibitor (50 units/l) 0.5 l

TransScript II RT 1 l


TransScript II Reverse Transcriptase[M-MLV,RNase H ]

(High Temperature RT)

Description

TransScript II Reverse Transcriptase is a recombinant M-MLV

reverse transcriptase with reduced RNase H activity and increased

thermostability. The enzyme is active up to 55oC. It provides higher

specicity, higher yield and more full-length cDNA products.

Increased thermostability for more full-length cDNA products.




cDNA up to 15 kb.Appl ication


Multiple copy and low-copy genes.

GC rich or comlex secondary-structure RNA template.

Unit Denition

One unit ofTransScript II is the amount of enzyme required to

incorporate 1 nmol of deoxyribonucleotide into acid-precipitable


primer .

Concentration

200 units/l

Components

TransScript II RT

10TS II RT Buffer (500 mM KCl; 30

mM MgCl2; 200 mM Tris-HCl pH 8.4)


Storageat -20oC for one year

AH101-02 10000 units


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1: REPA 1.8 kb; 2: NCBP 2.5 kb;

3: HDP 3.5 kb; 4: VIN 4.6 kb;

5: Pol 6.8 kb; 6: APC 8.5 kb;

7: Dynein 12.3 kb; 8: FAL 15.1 kb


M21 2 3 4 5 6 7 8M1

The condition for the PCR is the same as for EasyScript Reverse

Transcriptase(P37) .

AE301-02 50 rxns20 l system

AE301-03 100 rxns20 l sytecm

EasyScriptFirst-Strand cDNA Synthesis SuperMix

Kit Components

Description

EasyScript First-Strand cDNA Synthesis SuperMix provides

all the necessary components for cDNA synthesis from total or

poly(A)+ RNA. The first-strand cDNA products generated by the

supermix are suitable for cloning, PCR or qPCR applications.




cDNA up to 8 kb.

Appl ication


AE301-02 AE301-03

EasyScriptRT/RI Enzyme Mix 50 l 250 l

2ES Reaction Mix 500 l 2500 l

Random Primer (0.1 g/l) 50 l 100 l

Anchored Oligo(dT)18 Primer (0.5 g/l) 50 l 100 l

RNase-free Water 500 l 1 ml

Storage



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PROTOCOL

the First Strand cDNA synthesis1 Reaction Components

Component VolumeTotal RNA/mRNA Variable



or GSP 2 pmol

2ES Reaction Mix 10 l

EasyScriptRT/RI Enzyme Mix 1 l


2 Incubation


For random primer, incubate at 25oC for 10 min then at 42oC for 30

min.

3 Incubate at 85o

C for 5 min to inactive enzyme.RT-PCR

It is suggested to use 2-4 l reverse transcription products as PCR

templates.

The suggested reaction condition is the same as for EasyScript

Reverse Transcriptase ( P39 ).


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Kit Components

AT301-02 AT301-03

TransScriptRT/RI Enzyme Mix 50 l 250 l

2TS Reaction Mix 500 l 2500 l




PROTOCOL


Volume




or GSP 2 pmol

2TS Reaction Mix 10 l

TransScript RT/RI Enzyme Mix 1 l


2 Incubation


For random primer, incubate at 25

o

C for 10 min, then at 42

o

C for 30min.


RT-PCR


templates.


Reverse Transcriptase( P39 ).

AT301-02 50 rxns20 l system


TransScript First-Strand cDNA Synthesis SuperMix

Description

TransScript First-Strand cDNA Synthesis SuperMix provides all the

necessary components for cDNA synthesis from total or poly(A)+ RNA.

The rst-strand cDNA products generated by this supermix are suitable

for cloning, PCR or qPCR applications.




cDNA up to 12 kb.

Appl ication


Storage



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Description

Unique genomic DNA remover is combined with First-Strand cDNA

synthesis SuperMix to achieve simultaneous genomic DNA remove

and cDNA synthesis. After cDNA synthesis, gDNA remover and reverse

transcriptase are inactivated by heating at 85oC for 5 minutes.

Simultaneous genomic DNA removal and cDNA synthesis.

cDNA up to 12 kb.

Appl ication



2 Incubation


For random primer, incubate at 25oC for 10 min then at 42oC for 30

min.


RT-PCR


templates.



Kit Components

AT311-02 AT311-03

TransScript RT/RI Enzyme Mix 50 l 250 l

gDNA Remover 50 l 250 l

2TS Reaction Mix 500 l 2500 l


Anchored Oligo(dT)18Primer (0.5 g/l) 50 l 100 l


PROTOCOL

Volume


Anchored Oligo(dT)18 Primer (0.5 g/l) 1 l

or Random Primer (0.1 g/l) 1 l

or GSP 2 pmol

2TS Reaction Mix 10 l

TransScript RT/RI Enzyme Mix 1 l

gDNA Remover 1 l


TransScript One-Step gDNA Removal and cDNA

Synthesis SuperMixAT311-02 50 rxns20 l system

AT311-03 100 rxns20 l sytecm

Storage



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PROTOCOL


SuperMix for PCR

Storage


DescriptionTransScript All-in-One First-Strand cDNA Synthesis SuperMix for

PCR provides all the necessary components for cDNA synthesis from

total or poly(A)+ RNA. The SuperMix is provided at 5 concentration

and used at 1concentration by adding template and H2O. cDNA

generated with the SuperMix are suitable for regular PCR. The kit is

not suitable for qPCR.

1-tube format for simple and fast setup and reduce pipetting

variability.

Fast 30 min cDNA synthesis protocol.

cDNA up to 12 kb.

Kit Components

First Strand cDNA synthesis

1. Reaction Components

2. Incubate at 42oC for 30 min.

3. Incubate at 85oC for 5 min to inactive enzyme.

RT-PCR


templates.

The suggested reaction condition is the same as forEasyScript


AT321-01

5TransScriptAll-in-One SuperMix for PCR 200 l

RNase-free Water 1 ml

Volume

Total RNA/mRNA 50 ng-5 g/5-500 ng

5TransScriptAll-in-One SuperMix for PCR 4 l




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TransScript All-in-One First-Strand cDNA Synthesis

SuperMix for qPCR

Storage


DescriptionTransScript

All-in-One First-Strand cDNA Synthesis SuperMix for

qPCR provides all the necessary components for cDNA synthesis from

total or poly(A)+ RNA. The SuperMix is provided at 5 concentration

and used at 1concentration by adding template and H2O. cDNA

generated by the SuperMix are suitable for qPCR. The kit is not

suitable for regular PCR.

1-tube format for simple and fast setup and reduced pipetting

variability.

Fast 10 min cDNA synthesis protocol.

High compatibility with qPCR reagent.

Kit Components

First Strand cDNA synthesis1. Reaction Components

2. Incubate at 42oC for 10 min.

3. Incubate at 85oC for 5 sec to inactive enzyme.

qRT-PCRIt is suggested to use 2-4 l reverse transcription products as qPCR

templates.The suggested reaction condition is the same as forTransStartGreen

qPCR SuperMix ( P60 ).

AT331-01

5TransScriptAll-in-One SuperMix for qPCR 200 l

5TransScriptAll-in-One No-RT Control

SuperMix for qPCR20 l

RNase-free Water 1 ml

Volume


5TransScriptAll-in-One SuperMix for qPCR 4 l


AT331-01 50 rxns 20 l system

PROTOCOL


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AH301-03 100 rxns20 l sytecm

TransScript II First-Strand cDNA Synthesis SuperMix

Kit Components

DescriptionTransScriptII First-Strand cDNA Synthesis SuperMix provides all the

necessary components for cDNA synthesis from total or Poly(A)+ RNA.

It is an optimized cDNA synthesis kit withTransScriptII RT. First-Strand

cDNA can be synthesized at higher temperature resulting higher yield

and more full-length cDNA products.

Storage





cDNA up to 15 kb.

Appl icationFirst strand cDNA synthesis.

PROTOCOL


2. Incubation

For anchored oligo(dT)20primer or GSP, incubate at 50

o

C for 30 min. For random primer, incubate at 25oC for 5-10 min, then at 50oC for

30 min.

For GC-rich or complex secondary structure RNA template, incubate

at 55oC, 30 min.


Component Volume




or GSP 2 pmol

2TS II Reaction Mix 10 l

TransSciptIIRT/RI Enzyme Mix 1 l


RT-PCRIt is suggested to use 2-4 l reverse transcription products as PCR

templates.


Reverse Transcriptase ( P39 ).

Components AH301-02 AH301-03

TransScript II RT/RI Enzyme Mix 50 l 250 l

2TS II Reaction Mix 500 l 2500 l

Random Primer(N9) (0.1 g/l) 50 l 100 l




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AH311-02 50 rxns20 l systemAH311-03 100 rxns20 l system

TransScript II One-Step gDNA Removal and cDNASynthesis SuperMix

Description

Unique genomic DNA remover is combined with First-Strand cDNA

synthesis SuperMix to achieve simultaneous genomic DNA remove

and cDNA synthesis. After cDNA synthesis, gDNA remover and

reverse transcriptase are inactivated by heating at 85oC for 5 minutes.

Simultaneous genomic DNA removal and cDNA synthesis.

cDNA up to 15 kb.

Storage


Kit Components


2. Incubation


For random primer, incubate at 25oC for 5-10 min, then at 50oC for

30 min.


RT-PCR


templates.



PROTOCOL

Volume


Anchored Oligo(dT)20 Primer (0.5 g/l) 1 l

or Random Primer (0.1 g/l) 1 l

or GSP 2 pmol

2TS II Reaction Mix 10 l

TransScript II RT/RI Enzyme Mix 1 l

gDNA Remover 1 l


Components AH311-02 AH311-03

TransScript II RT/RI Enzyme Mix 50 l 250 l

gDNA Remover 50 l 250 l

2TS II Reaction Mi

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