bioneer catalog 2006 - Bioneer Catalog 2006.pdf · Enzymes Enzymes 98 Fig 2. Extension time was limited to 60 sec and 1 ~ 10 kb of lambda DNA target gene was amplified. Bioneerss

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6. Enzymes

Polymerases & Modifying Enzymes

Restriction Enzymes

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Polymerases & Modifying Enzymes

Taq DNA Polymerase

Product Size Contents Cat No.

Description: The Polymerase gene from Thermus aquaticus

(strain YT1) is cloned and expressed in E. coli, then Taq DNA

polymerase is produced with high purity. It is used in the

amplification and sequence testing of DNA through PCR. The

quality of the Taq DNA polymerase has been tested by the Activity

Test, SDS-Page, Endonuclease Test, etc.

Supplied with Enzyme

10 x Reaction buffer: 100 mM Tris-HCl, 400 mM KCl, 15 mM

MgCl2, pH 9.0.

Dilution Buffer: 20 mM Tris-HCl, 100 mM KCl, 0.5 mM EDTA,

1 mM DTT, 0.5 % Tween 20, 0.5 % IGEPAL CA-630 (pH 8.0.)

Storage Condition

20 mM Tris-HCl, 100 mM KCl, 0.5 mM EDTA, 1 mM DTT, 0.5 %

Tween 20, 0.5 % IGEPAL CA-630 (pH 8.0.) Store at -20

Concentration: 5 units/

Unit Definition

One unit is defined as the amount of enzyme required to catalyze

the incorporation of 10 nmole of dNTPs into an acid-insoluble form

in 30 minutes at 72

Activity Test

Working Range Test & Sensitivity Test

Using 1 unit of Taq DNA polymerase, the activity of the polymerase

was tested on human, bacterial, and lambda genomic DNA as

template. Each template DNA was serially diluted by tenfolds, with

different ranges.

A: Lambda genomic DNA (100 ng ~10 fg)

B: Bacteria genomic DNA (100 ng ~10 fg)

C: Human genomic DNA (100 ng ~10 pg)

Fig 1. Test of working range & sensitivity of Taq DNA

polymerase for each template. The PCR mixture was mixed with

serially diluted template DNA and 20 pmole of Forward/Reverse

primers, dNTPs Mix, and 10 x reaction buffer; diluted Taq DNA

polymerase was added just before PCR was carried out. From 10

fg to 100 ng of lambda genomic DNA was amplified, 10 fg to 100

ng of bacterial genomic DNA was amplified, and 10 pg to 100 ng of

human genomic DNA was amplified.

Lane 1: 100 ng of template DNA Lane 2: 10 ng of template DNA



Lane 7: 100 fg of template DNA Lane 8: 10 fg of template DNA

Examination of Taq DNA Polymerase working speed

The working speed of Taq DNA Polymerase wae tested by limiting

the extension time to 60 seconds. While conventional Taq DNA

polymerases had an average speed of ~ 60 nts/sec.

Enzyme Optimum

temp.( )

Half life(min, in

95 )

MgCl2(mM)

KCl(mM)

Optimum

pH (25 )

Taq 72 80 1.5 40 8.3

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

A B C

Taq DNA Polymerase 250 units 10 mM dNTPs, 10 x reaction buffer with MgCl2 E-2010



Taq DNA Polymerase 250 units 10 mM dNTPs, 10 x reaction buffer free MgCl2 , 20 mM MgCl2 E-2010-1

Taq DNA Polymerase 500 units 10 mM dNTPs, 10 x reaction buffer free MgCl2, 20 mM MgCl2 E-2011-1

Taq DNA Polymerase 1000 units 10 mM dNTPs, 10 x reaction buffer free MgCl2, 20 mM MgCl2 E-2012-1

Taq DNA Polymerase 250 units 10 x reaction buffer with MgCl2 E-2010-2



Taq DNA Polymerase 250 units 10 x reaction buffer free MgCl2, 20 mM MgCl2 E-2010-3



dNTP 1 ml 10 mM ( each 2.5 mM ) D-3001

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Fig 2. Extension time was limited to 60 sec and 1 ~ 10 kb of

lambda DNA target gene was amplified. Bioneer s Taq DNA

Polymerase amplified up to 10 kb of the target gene within the

60 sec extension period.

Lane M: 1 Kb DNA Ladder (D-1040, Bioneer)

Lane 1: 1 kb PCR Product Lane 2: 2 kb PCR Product





Efficiency test of Taq DNA polymerase

The amplification efficiency was examined according to the number

of cycles. PCR product after 30, 40, 50, 60 cycles was quantified

by fluorometer (Phamacia corp.) and the efficiency was determined

by the equation below 1 ng of human genomic DNA used.

Fig 3. M: 100 bp DNA Ladder (D-1030, Bioneer)

Lane 1: 30 cycle, Lane 2: 40 cycle, Lane 3: 50 cycle, Lane 4: 60

cycle

Nf= No (1+Y)n

- Nf: product copies following n cycles

- No: no. of template copies

- Y: efficiency of Taq DNA polymerase

- n: no. of PCR cycles

Reliability Test

To test the reproducibility of the produced batch, the Taq DNA

polymerase from each lot was serially diluted and the range of

amplification tested

Fig 4. Reliability Test PCR was carried out with Taq DNA

polymerase serially diluted to 1/5, 1/10, 1/15, 1/20 on lambda (A)

and human (B) genomic DNA as template. DNA carried out. The

amplification range of each lot is equal.

Lane 1: 5 U of Taq DNA pol. Lane 2: 1 U of Taq DNA pol.

Lane 3: 0.5 U of Taq DNA pol. Lane 4: 0.33 U of Taq DNA pol.

Lane 5: 0.25 U of Taq DNA pol.

Related Product

AccuPower TM PCR PreMix

AccuPower TM HL PCR PreMix

AccuPower TM RT PreMix

AccuPower TM RT/PCR PreMix

* This product is sold under licensing arrangements with Roche Molecular Systems, F.

Hoffmann-La Roche Ltd ( Roche ) and Perkin-Elmer Corporation. Purchase of this

product is accompanied by a license to use it in the Polymerase Chain Reaction

(PCR) process in conjunction with an Authorized Thermal Cycler.

<A> <B>

M 1 2 3 4

1 2 3 4 5 1 2 3 4 5

2.50E+12

2.00E+12

1.50E+12

1.00E+12

5.00E+11

0.00E+0020 30 40 50 60 70

Cycle No.

No. of Cycles Copy No. Efficiency of Taq DNA Pol.

30 6.20 E + 11 104%

40 1.55 E + 12 75%

50 1.99 E + 12 57%

60 2.08 E + 12 46%

M 1 2 3 4 5 6 7 8 9 10

Co

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Description: HL Taq DNA Polymerase is themostable DNA

polymerase for high fidelity, long and high yield PCR. Also, it has

heat-stability, convenience, simplicity and reproducibility.

Feature & Benefits

HL Taq DNA polymerase is a unique enzyme containing

mixture of Taq DNA polymerase and the thermostable DNA

polymerase with proofreading activity. It suits well not only for

conventional PCR but also for the amplification of large DNA

template. The HL Taq DNA polymerase allows amplification of

DNA fragment up to 20 kb from lambda DNA.

HL Taq DNA polymerase offers about three times the fidelity

(1.89x10-6) and eight times the yield of other Taq DNA

polymerase.


10 x Reaction Buffer (1ml): 100 mM Tris-HCl, 400 mM KCl, 15

mM MgCl2, pH 9.0

Dilution Buffer (1ml): 20 mM Tris-HCl, 0.5 mM EDTA, 1 mM

DTT,100 mM KCl, Stablizers, 50 % Glycerol, pH 8.0

dNTPs mixture (0.5 ml): 10 mM, each dNTP 2.5 mM

Storage Condition

20 mM Tris-HCl, 0.5 mM EDTA, 1 mM DTT, 100 mM KCl,

Stablizers, 50 % Glycerol pH 8.0, store at -20


Unit Definition

One unit is defined at the amount of enzyme that will incorporates 10

nmol of dNTPs into acid-insoluble material in 30 minutes at 72 .

Quality Assurance

Nuclease activity is not detected after incubation of 1 ug of

substrate DNA-supercoiled plasmid and lambda/Hind III DNA - with

5 units of HL Taq DNA Polymerase in 50 uL reaction volume with

the supplied reaction buffer for 18 hrs at 37 and 70 .

General Reaction Condition [ 20 reaction volume ]

Reaction mixture

Template variable

Primer (forward) 5 ~ 10 pmoles

Primer (reverse) 5 ~ 10 pmoles

10 x Reaction buffer 2

10mM dNTPs mix. variable

Taq DNA Polymerase 0.5 ~ 1.0 unit

D.W variable

---------------------------------------------------------

Total 20

Note

In extremely long PCR, we recommend designing primers 35 ~

40 length bases containing 40 ~ 60% G+C residues with Tm of

68 ~ 72 and performing two step PCR - 94 for 30 sec, 68

for 1 min / 1 kb, 30 rounds.

Conventional

PCR (< 10kb)

DNA : 1 ~ 20 ngTemplate 5 ~ 50 ng Bacteria : 20 ~ 100 ng

Human : 100 ~ 500 ng

Primer 10 ~ 30 pmoles 5 ~ 20 pmoles

Extremely long PCR

(10 kb ~ 20 kb)

HL Taq DNA Polymerase

HLTaq DNA Polymerase 250 Units Each E-2018

HLTaq DNA Polymerase 1,000 Units Each E-2019

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HotStart Taq DNA Polymerase


Description

HotStart Taq DNA Polymerase is a modified form of the recombinant

Taq DNA polymerase. The activity of HotStart Taq DNA polymerase

is completely inhibited when the temperature is below 70 . The

enzyme requires an activation step at 95 for 10~15 minutes . The

thermal activation prevents the extension of non-specifically

annealed primers and primer-dimers formed at low temperature

during PCR setup. The reaction condition of HotStart Taq DNA

polymerase can also be used without any modification for any PCR

reaction condition and cycling parameter of Taq DNA polymerase.

Activation step

HotStart Taq DNA polymerase needs to be activated for 15

minutes at 95 as initial step of PCR

Features and BenefitsHigh Specificity

High Sensitivity

Improve product yields

Reduce nonspecific background

Easy handling (Reliable Room-temperature setup)

ApplicationHotstart PCR

Low copy numbers PCR

Multiplex PCR

Quantitative PCR

Unit Definition

One unit is defined as the amount of enzyme required to catalyze

the incorporation of 10nmoles of dNTP into acid-insoluble material

in 30 minutes at 72 .

Quality Assurance

Nuclease activity is not detected after incubation of 1 ug of substrate

DNA - supercoiled plasmid and lambda/Hind III DNA - with 5 units

of HotStart Taq DNA Polymerase in 50 uL reaction volume with the

supplied Reaction buffer for 18 hr at 37 oC and 70 .

Supplied with10X Reaction Buffer : Contains Tris-HCl, KCl, 15 mM MgCl2, pH

9.0

dNTPs mixture : 10 mM, The concentration of each dNTPs is 2.5

mM.

Dilution and storage buffer : 20 mM Tris-HCl, 0.5 mM EDTA, 1

mM DTT, 100 mM KCl, Stabilizers, 50 % Glycerol, pH 8.0

Storage condition

HotStart Taq DNA polymerase, including buffers and reagents,

should be stored immediately upon receipt at -20 oC in a constant

temperature freezer.

Activity Test

Sensitivity test

The result of sensitivity comparison between One of the well-

known competitive companies (Q)’s HotStart and Bioneer’s

HotStart Taq DNA polymerase using Lambda and Human

Genomic DNA as template is shown below. 1 U of each enzymes

were used per reaction and each template DNA is continuously

diluted 10 times.

HotStart Taq DNA polymerase Competitive (Q) Hotstart Product

Fig 1. Sensitivity test Compared with Bioneer’s HotStart Taq

DNA polymerase and Qiagen’s Hotstart product. A) Using

Lambda DNA as a template, they were amplified up to 1kb

fragment length by having 5X10^8 copies to 5 copies B) Using

Human DNA as a template, they were amplified P53 gene of 400

bp fragment by having 5X10^4 copies to 50 copies. Lane 6 is

template negative.

Specificity test

Using Human genomic DNA (10ng) as a template, specificity

comparison is done between Taq DNA polymerase, HotStart Taq

DNA polymerase and the well-known Competitor company (Q)°Øs

HotStart and is confirmed specificity by amplifying P53 gene up to

1560bp (from 140bp to 1560) by using primer pairs that is over

65% GC content and 10?C Tm differences between each forward

and reverse primer.

HotStartTaq DNA Polymerase 250 Units 10mM dNTPs, 10 reaction buffer with MgCl2 E-2017

HotStartTaq DNA Polymerase 1000 Units 10mM dNTPs, 10 reaction buffer with MgCl2 E-2017-1

M 1 2 3 4 5 6 7 8 9 10

M 1 2 3 4 5 6 M 1 2 3 4 5 6

A)

B)

M 1 2 3 4 5 6 7 8 9 10

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Taq pol. HotStart Taq pol. Competitor CO.(Q)HotStart

Fig 2. Specificity Comparison between Taq, HotStart Taq DNA

Polymerase and Competitor company (Q)’s HotStart.

Enzyme dilution test

Amplify Human P53 of 600bp fragment after having HotStart Taq

DNA polymerase repeatedly diluted in the same sequential units

from 2U up to 0.125U(2U:1:0.5:0:25:0.125).

Fig 3: The amplification result of P53 gene (600bp) from Human

genomic DNA per units of HotStart Taq DNA polymerase.

M: 100bp DNA Ladder

Line 1: Enzyme 2 U Line 2: Enzyme 1 U

Line 3: Enzyme 0.5 U Line 4: Enzyme 0.25 U

Line 5: Enzyme 0.125 U Line 6: Enzyme negative

Multiplex PCR

1) Single & multiplex PCR using one template and five primer set.

Fig 4 : Result of Single & Multiplex PCR. Amplified P53 Gene from

Human Genomic DNA. The result of specific primer set is used in

each different lane from 1 to 5 and PCR reaction in one tube of the

primer set mixtures of lane from 1 to 5 in lane 7.

M : 100bp DNA Ladder

Line 1: 119bp fragment Line 2: 246bp fragment


Line 5: 600bp fragment Line 6: Template DNA negative

Line 7: Multiplex PCR with primer mixtures from 1 to lane 5

2) Proceeded PCR reaction from one tube using 6 sets of primer

and 6 different types multiplex PCR of genomic DNA from six

different primer-template system.

HotStart Taq pol. HotStart Product (Company Q)

Fig 5 : Result of Single & Multiplex PCR.. From lane 1 to 6 is the

result of PCR reaction of genomic DNA and specific primer. Lane 7

is the PCR reaction in one tube of the primer set mixtures of lane

from 1 to 6. Lane 8 is the Competitor company(Q)’s multiplex PCR

kit..

M: 100bp DNA Ladder




Line 7: Multiplex PCR with template DNA & primer mixtures from 1

to lane 6

Quantitative PCR

Real Time PCR Comparison data chart Using HotStart Taq DNA

polymerase and Company (Q)’s HotStart product. The result of

DNA Titration From 5 copies to 5x10^8 copies are as follows:

efficiency=0.30, Linearity = 0.999

HotStart Taq pol. HotStart Product (Company Q)

Fig 7. Quantitative PCR comparison between HotStart Taq-pol

and Company (Q)’s HotStart

Ordering Information

CAT.#

E-2017

E-2017-1

PRODUCT NAME

HotStart Taq DNA polymerase, 250Unit, 10 mM

dNTPs, 10 X reaction buffer with MgCl2

HotStart Taq DNA polymerase, 1000Unit, 10 mM

dNTPs, 10 X reaction buffer with MgCl2

M 1 2 3 4 5 6 7

M 1 2 3 4 5 6

M 1 2 3 4 5 6 7

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

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Pfu DNA Polymerase


Description : Pfu DNA polymerase is a thermostable DNA

polymerase isolated from Pyrococcus furiosus Vc1. It catalyzes the

DNA-dependent polymerization of nucleotides into duplex DNA in

the 5 3 direction and exhibits 3 5 exonuclease (proof

reading) activity. Pfu DNA polymerase is the ideal choice for a

variety of techniques requiring high-fidelity DNA synthesis by PCR

reaction. It can apply to cloning, gene expression, site-directed

mutagenesis and etc.

Features and Benefits

High-Fidelity: 3 - 5 exonuclease(proofreading) activity

Thermostability: retaining 94-99% of its thermostable

activity after 1 hour at 95

Modified Nucleotide incorporation: incorporates the following

modified nucleotides: 7-deaza-dGTP, a-thionucleotide,

fluoresceinated and biotinylated nucleotides.

Terminal Transferase Activity: devoid of terminal transferase

activity and generates blunt-ended PCR products.

Applications

- Polymerase Chain Reaction (PCR)

- Primer Extension

- Gene Cloning

- Gene Expression

- Site-directed Mutagenesis


10 x Reaction Buffer (1 m ): 200 mM Tris-HCl, 100 mM KCl,

100 mM (NH4)2SO4, 20 mM MgSO4, 1% Triton x -100, 1 mg/ml

Acetylated BSA, pH 8.8

Storage Condition

50 mM Tris-HCl, 0.1 mM EDTA, 1 mM DTT, Stablizers, 50%

Glycerol, pH 8.2, store at -20

Concentration : 2.5 units/

Unit Definition

One unit is defined at the amount of enzyme that will incorporate 10

nmol of dNTP into acid-insoluble material in 30 minutes at 72

Quality Assurance

Nuclease activity is not detected after incubation of 1 ug of

substrate DNA-supercoiled plasmid and lambda/Hind III DNA- with

5 units of Pfu DNA Polymerase in 50 uL reaction volume with the

supplied 10 reaction buffer for 18 hrs at 37 and 70 .

Activity Test

Amplication of limiting amounts of template DNA: comparison of

Lambda DNA (A), Bacterial genomic DNA (B), Human genomic

DNA (C).

* Lambda DNA was amplified using 2.5 units of enzyme in 50 uL reaction volume.

M: 1 Kb DNA Ladder (D-1040, Bioneer)

Lane 1: 10 ng of Lambda DNA

Lane 2: 1 ng of Lambda DNA

Lane 3: 100 pg of Lambda DNA



Lane 6: 100 fg of Lambda DNA

Lane 7: 10 fg of Lambda DNA

* Bactierial DNA was amplified using 2.5 units of enzyme in 50 uL reaction volume.


Lane 1: 10 ng of Bacterial DNA

Lane 2: 1 ng of Bacterial DNA

Lane 3: 100 pg of Bacterial DNA



Lane 6: 100 fg of Bacterial DNA

* Human DNA was amplified using 2.5 units of enzyme in 50 uL reaction volume.


Lane 1: 10 ng of Human DNA

Lane 2: 1 ng of Human DNA

Lane 3: 100 pg of Human DNA



Lane 6: 100 fg of Bacterial DNA

Amplification of fragments ranging from 500 bp to 5 kb from

template Lambda DNA 100 pg with 2.5 units of Pfu DNA

Polymerase.

M: 1 Kb DNA Ladder

(D-1040, Bioneer)

Lane 1: amplification of 500 bp

fragment

Lane 2: amplification of 1.0 Kb

fragment

Lane 3: amplification of 2.0 Kb

fragment

Lane 4: amplification of 3.5 kb fragment

Lane 5: amplification of 5.0 kb fragment

M 1 2 3 4 5 6 7

M 1 2 3 4 5 6

M 1 2 3 4 5

A

B

C

Pfu DNA Polymerase 250 Units 10 x reaction buffer, without dNTPs E-2015

Pfu DNA Polymerase 1,000 Units 10 x reaction buffer, without dNTPs E-2016

dNTP 1 ml 10 mM ( each dNTPs, 2.5 mM ) D-3001

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M-MLV Reverse Transcriptase


T7 RNA Polymerase

Description : Moloney Murine Leukemia Virus (M-MLV) Reverse

Transcriptase is an RNA-dependent DNA polymerase. This

enzyme is able to use an RNA molecule as a template and

synthesize a double-strand DNA .

Source

M-MLV Reverse Transcriptase is isolated from a E.coli strain

containing a recombinant clone.

Applications

First-strand synthesis of cDNA from RNA molecules.


5 x Reaction Buffer (0.5 ml): 250 mM Tris-HCl, 150 mM KCl,

40 mM MgCl2, pH 8.3

100 mM DTT: 0.3 ml

Storage Condition

20 mM Tris-Cl (pH7.6), 150 mM NaCl, 0.1 mM EDTA, 1 mM DTT,

0.1% IGEPAL CA-630, 50% Glycerol, store at -20


Unit Definition

One unit is defined as the amount of enzyme required to

incorporate 1 nmole of dTTP into acid-precipitable material in 10

min at 37 using oligo d (T) as template primer.

Quality Assurance

DNase and RNase activity is not detected after incubation of 1ug of

DNA and RNA with 200 units of M-MLV Reverse Transcriptase for

3 hours at 37 ~ 42 .

References

1. Gerard, G. F. et al. (1975) J. Virol. 15, 785 - 797

2. Roth, M. et al. (1985) J. Biol. Chem. 260, 9326 - 9335

3. Sambrook, J. Fritsch, E. F. and Maniatis, T. (1989) Molecular

Cloning: A Laboratory Manual, second edition, Cold Spring

Harbor, New York, 8. 64.

Related Products

AccuPower TM RT PreMix

AccuPower TM RT/PCR PreMix

AccuRapid TM PCR cDNA Library Kit


Description: T7 RNA Polymerase is a DNA dependent RNA

polymerase with a highly specificity for the initiation of transcription

at T7 RNA polymerase promoters. It is widely used for the rapid

synthesis in vitro of specific RNAs.

Source

T7 RNA Polymerase is isolated from E.coli cells containing the

ligase gene cloned from T7 bacteriophage.

Applications

Synthesis of RNA transcripts for northern hybridization and

southern hybridization probes.

RNA generation for studies of RNA structure, procession and

catalysis


10 x Reaction Buffer (1 ml): 400 mM Tris-Cl (pH 8.0), 60 mM

MgCl2, 20 mM Spermidine

100 mM DTT (0.5 ml)

DEPC-DW (1ml)

Storage Condition

200 mM Na-phosphate (pH 7.7), 10 mM NaCl, 1 mM EDTA, 1 mM

DTT, 0.02 % Triton x -100, 0.08% Tween-20, 50% Glycerol, Store

at -20

Concentration: 0.1 units/

Unit Definition

One unit of enzyme catalyzed incoporation of 1 nmoles of

[3H]ATPs into acid insoluble form in 60 min at 37

Quality Assurance

Nuclease Contamination Assay: No altered was detected after

incubation of 1 ug of substrate DNA with 500 units of T7 RNA

polymerase for 18 hrs in 37

Protease Contamination Assay: No altered pattern was

detected after in cubation of 2,000 units of T7 RNA polymerase

for 18 hrs in 37

M-MLV Reverse Transcriptase 10,000 Units 5 x reaction buffer, 0.5 ml, 100 mM DTT, 0.3 ml E-3121

M-MLV Reverse Transcriptase 50,000 Units 5 x reaction buffer, 2.5 ml, 100 mM DTT, 1.5 ml E-3122

T7 RNA Polymerase 2,000 Units 10 x reaction buffer, 1 ml, 100 mM DTT, 0.5 ml, DEPC-DW, 1 ml E-3041

T7 RNA Polymerase 10,000 Units 10 x reaction buffer, 5 ml, 100 mM DTT, 2.5 ml , DEPC-DW, 5 ml E-3042

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DNA Polymerase 1, Large (klenow) Fragment


Description

DNA Polymerase I, Large (Klenow) Fragment is derived from E.coli

DNA polymerase that it has the function of Polymerization an 3’ 5’

exonuclease but it has deficient activity of 5’ 3’ exonuclease.

Source

DNA Polymerase I, Large (Klenow) Fragment is isolated from an E.coli

strain carrying the gene of the enzyme on a plasmid.

Applications

DNA sequencing by the dideoxy method

Random priming labeling

Second strand synthesis in oligonucleotide directed mutagenesis

Reaction Condition

10 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 1 mM DTT, Incubate at 37 .

Storage Conditions

20 mM Tris-HCl (pH 7.5), 50 mM KCl, 1 mM EDTA, 10 mM 2-

mercaptoethanol, 50% Glycerol,

Store at -20

Unit Definition

One unit of enzyme is defined as the amount of enzyme required to

convert 10 nmoles of dNTPs to an acid-soluble form in 30 min at 37 .

Quality Assurance

Nuclease Contamination Assay: No altered was detected after

incubation of 1ug of substrate DNA with 100units of DNA

Polymerase I (Klenow Fragment) for 18 hrs in 37 .

Protease Contamination Assay: No altered pattern was detected

after incubation of 100 units of DNA polymerase I (Klenow Fragment)

for 18 hrs in 37 .

Concentration : 5 units/

DNA Polymerase I 200 U Each E-3021DNA Polymerase I 1,000 U Each E-3022

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Description: T4 DNA Ligase catalyzes the formation of a

phosphodiester bond between juxtaposed 5 phosphate and 3

hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt-

end and cohesive-end termini as well as repair single stranded

nicks in duplex DNA, RNA, or DNA/RNA hybrids.

Source: T4 DNA Ligase is isolated from a recombinant E.coli

strain.

Applications: Joining double-stranded DNA with cohesive or

blunt ends


10 x Reaction Buffer (1 ml) : 500mM Tris-HCl (pH 7.8),100mM

MgCl2, 50mM DTT, 10mM ATP, 25 ug/ml BSA

Storage Condition

10mM Tris-HCl (pH 7.5), 50mM KCl, 1mM EDTA, 10 mM 2-

mercaptoethanol, 50% glycerol, store at -20 .

Concentration: 1 unit /

Unit Definition: 0.01 Weiss unit of enzyme is defined as the

amount of enzyme required to give 90% ligation of Hind lll

fragments of lambda DNA in 30 min at 16 in 20 of the assay

mixture.

Quality Assaurance: No altered was detected after incubation of

1 substrate DNA with 10 units of T4 DNA Ligase in 20 ul reaction

volume with the supplied reaction buffer for 18 hrs at 37

Note Store the buffer in small aliquots at -20 to minimize

degradation of the ATP and DTT

Ligation Test


Lane 1: DNA fragment (digested with EcoR V)

Lane 2,3,4,5: T4 DNA Ligase 1 unit, 16 , 10 min, 20 min, 30 min, 60 min



Lane 14: lambda DNA (digested with Hind III)

Lane 15,16,17: T4 DNA Ligase 0.01 unit , 16 , 10 min, 20 min, 30 min



Blunt-end ligation Cohesive-end ligation

1 2 3 4 5 6 7 8 9 10 12 13 14 15 16 17 18 19 20 21 22 23 24

T4 DNA Ligase

T4 DNA Ligase 100 Weiss unit Each E-3061

T4 DNA Ligase 500 Weiss unit Each E-3062

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Thermostable Thermus Filiformis (Tfi) DNA Ligase


Description : Tfi DNA Ligase plays role of formatting

phosphodiester bond by connecting double stranded DNA or

connecting oligonucleotides 3’-hydroxyl end and 5’-phosphate end

together. Especially the reaction temperatures are between 45

and 65 compared to DNA Ligases such as T4 DNA Ligse, E.coli

DNA Ligase, etc. It is the higher temperature that keeps safe and

active. In addition, Tfi DNA Ligase is possible to use in ligation

reaction in high reaction temperature requirement.

Source

Tfi DNA Ligase is isolated from E.coli cells containing the ligase

gene cloned from Thermus filiformis.

Applications

Ligase Chain Reaction (LCR)

Oligonucleotide Ligation Assay (OLA)

Mutagenesis by Incorporation of a phosphorylated oligo during

PCR Amplication

Simultaneous Mutagenesis of Multiple Sites


10 x Reaction Buffer (1 ml): 300 mM Tris-HCl (pH8.3), 250 mM

KCl, 50 mM MgCl2, 5 mM NAD

1 x Dilution Buffer (1 ml): 10 mM Tris-HCl (pH7.6), 0.1 mM

EDTA, 50 mM KCl, 1 mM DTT, 200 ug/mL acetylated BSA,

50% Glycerol

Storage Condition

20 mM Tris-HCl (pH7.6), 2 mM MgCl2, 1 mM EDTA, 1 mM DTT,

0.5% Tween -20, 0.5% IGEPAL CA-630, 50% Glycerol, store at -

20 .

Concentration : 20 units/

Unit Definltion

One unit of Tfi DNA Ligase is defined as the amount of enzyme

required to give 50% ligation of the 12 base pair cohesive ends of 1

ug of PspEI digested lambda DNA in 10min at 45 .

Activity Assay Conditions

The activity assay is carried out in a 20 reaction containing 1 ug

of PspEI digested lambda DNA and 1 x Tfi DNA ligase reaction

buffer. After incubation at 45 for 10 min., the reaction is

terminated by addition of stop solution (40%(w/v) sucrose, 50 mM

EDTA and 0.25% bromophenol blue). Then heat at 70 for 10

min. and immediately load on a 0.8% agarose gel.

Stability

The half-life of the enzyme in 1 x reaction buffer is more than 1

hour at 95 and 55 hours at 65 .

Note : Tfi DNA Ligase should not be used as a substitute for other

DNA ligases, i.e., T4 DNA Ligase.

References

1. Barany, F. (1991) Proc. Natl. Acad. Sci. USA, 88, 189 - 193.

2. Landegren, U. et al.(1988) Science 241, 1077 - 1080

3. Michael, Scott F. (1994) Biotechniques 16:3, 410 - 412.

4.Gerard J. A. et al. (1993) Biotechniques 15:1, 172 - 178.

1 2 3 4 5 6

LigationProduct(1+4)

1. Fragment

4. Fragment

Lane 1: DNA/PspE I (control)

Lane 2: Incubate at 45 , 10 min




Lnne 6: Incubate at 65 , 10 min

Control Condition: Incubate the reaction containing ligase 1 unit and

1 ug DNA[lambda PspEI] at each temperature for 10 min.

Test of temperature for ligation (45 ~ 65 )

Tfi DNA Ligase 2,000 Units 10 x reaction buffer, 1 ml, 1 x dilution buffer, 1 ml E-3111

Tfi DNA Ligase 10,000 Units 10 x reaction buffer, 5 ml, 1 x dilution buffer, 5 ml E-3112

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1 2 3 4 5 6 7 8 9 10

Lane 1: DNA/PspE I (control) Lane 2: Incubate at 95 , 10 min

Lane 3: Incubate at 95 , 20 min Lane 4: Incubate at 95 , 30 min




Control condition : Incubate the enzyme at 95 each time. Andthen add 1unit ligase to a 20 reaction containing 1ugDNA[lambda PspEI] and incubate the mixture at 45 for 10 min.

1 2 3 4 5 6 7 8 9 10 11 12 13 14

Lane 1: DNA/PspE I (control) Lane 2: Incubate at 65 , 6 hrs

Lane 3: Incubate at 65 , 12 hrs Lane 4: Incubate at 65 , 18 hrs






Control condition: Incubate the enzyme for each time at 65

and then add 1 unit ligase to a 20 reaction containing

1 ug DNA[lambda PspEI] and incubate the mixture at 45 for 10

min.

Heat stability test at 95 and 65

Enzymes

En

zym

es

108

Description

Tte Inorganic Pyrophosphatase (Ppase) is enzyme to make

orthophosphate form by hydrolyzing Inorganic Pyrophosphatase.:

P2O7-4+H2O -> 2HPO4

-2

Source

Tte Inorganic Pyrophosphatase (PPase) is isolated from Thermus

Thermophilus B35.

Storage Buffer

20 mM Tris-HCl (pH 8.0), 0.2 % Triton X-20, 100 mM KCl, 0.1 mM

EDTA,

50 % Glycerol, store at -20

Unit Definition

One unit is the amount of enzyme that will generate 40nmoles of

phosphate per minute from pyrophosphate under standard

reaction conditions (50 mM Tris-HCl (pH8.5), 1 mM MgCl2, 0.32

mM PPi) at 65 .

Concentration

300 units/ml

Reference : Heinonen, J. K, and Lahti, R. J. (1981) Analytical

Biochemistry 113, 313-317

Product Unit Contents Cat No.

PPase 0.5 Each E-3071

PPase 2.5 Each E-3072

Tte Inorganic pyrophophatase(PPase)

BstSE(Nickase) GAGTC (4/-)

Description

BstSE is site specific endonuclease that inltiates cleavage only in

single strand of double strand DNA

Source

BstSE is isolated from Bacillus Stearothermophilus SE-589.

Reaction Buffer

10 mM Tris-HCl (pH 8.5), 10 mM MgCl2, 150 mM KCl, 1 mM DTT,

Incubate at 55 .

Storage Conditions

10 mM Tris-HCl (pH 7.5), 50 mM KCl, 1 mM EDTA, 10 mM 2-

mercaptoethanol, 50 % Glycerol, store at -20

Ligation and Recutting

After 5-fold overdigestion with BstSE, 90 % of the DNA fragments

can be ligated and recut.

Concentration

5 units/

Note : BstSE catalyzes a single-strand nick on the 3 side of the

recognition sequnece.


BstSE (Nickase) GAGTC (4/-) 50 Each E-2181

BstSE (Nickase) GAGTC (4/-) 250 Each E-2182

Enzymes

En

zyme

s

109

Bovine Serum Albumin(BSA), Acetylated

Description

Acetylated BSA is an enzyme stabilizer that is essential carrier

protein to remove contaminants such as nuclease and protease.

Acetylation process does not affect specific character of BSA nor

PCR but it removes nuclease’s activation very accurately. Add

BSA to be 5~200 /ml as a final concentration.

Source Bos Taurus

Storage Conditions

10 mM Tris-HCl (pH 7.5), Store at -20

Concentration

20 mg/ml

Reference

Gonzales, N., Wiggs, J., and Chamberlin, M. J. (1977) Arch.

Biochem. Biophys. 182, 404 - 408


BSA 20 mg Each E-3091

BSA 100 mg Each E-3092

Enzymes

En

zym

es

110

AccuRapidTM DNA Ligase

Description

AccuRapidTM DNA Ligase is a product can ligate cohesive-ended

DNA fragment and blunt-ended DNA fragment within 5 minutes

that reaction optimal temperature is from 15 to 25 .

AccuRapidTM DNA Ligase Buffer is the buffer type of product that

can give high efficient ligation result through cohesive-end and

blunt-end DNA fragment reaction. Ligation product is possible to

use directly into transformation.

Cat No. K-7101 Contents: For 50 ligations

50 unit(1 unit/ )

AccuRapidTM DNA Ligase(from T4 bacteriophage)

0.5 ml

2x AccuRapidTM DNA ligation buffer

Cat No. K-7102 Contents : For 150 ligations

50 unit x 3 (1 unit/ )

AccuRapidTM DNA Ligase(from T4 bacteriophage)

0.5 ml x 3

2x AccuRapidTM DNA ligation buffer

Storage Condition : Store at -20

Quality Control Assay: The kit is pre-tested in a ligation

according to the standard protocol; 500 ng of blunt-ended 500 bp

insert DNA was ligated to 100 ng of pUC19 DNA digested with

Sma I. Vector DNA fragments should most preferably be

dephosphorylated to minimize self-circularization. Yield of white

colonies after transformation is > 90%

Quality Assaurance: Nuclease activity is not detected after

incubation of 1 of substrate DNA with 10 units of AccuRapid

DNA Ligase in 20 reaction volume with the supplied Reaction

buffer for 18 hrs at 37

Reference

feiffer, B. H. and Zimmerman, S,B., Nucleic Acids Research, 11,

7853 ~ 7871, 1983.

Hayashi., Nakazawa, M., Ishizaki, Y., Hiraoka, N., and Obayashi,

A., Nucleic Acids Research, 13, 7979 ~ 7992, 1985.

Ligation test of reaction temperatures

Lane 1: 1 Kb DNA Ladder(D-1040)

Lane 2: EcoR V digested DNA fragments

Lane 3: Ligation reaction with T4 DNA Ligase reaction

buffer(E-3061) at 37 for 5 min

Lane 4: Ligation reaction with AccuRapid DNA Ligation

Kit at 16 for 5 min


Kit at 25 for 5 min


Kit at 37 for 5 min

Product Size Cat No.

AccuRapidTM DNA Ligation Kit 50 ligations K-7101

AccuRapidTM DNA Ligation Kit 150 ligations K-7102

Enzymes

En

zyme

s

111

B

B

I

B

O

I

I

N

I

B

I

N

B

I

N

R

B

I

R

N

O

R

B

R

R

O

O

B

I

O

O

N

R

O

R

Bsp13 I

B

O

I

B

B

I

O

I

B

B

B

O

I

I

I

I

Acc16 I

Acc65 I

Acc113 I

AccB1 I

AccB7 I

AccBS I

Acl I

AclN I

AclWI

Acs I

Afe I

Alu I

Ama87 I

Apa I

AsiA I

AspLE I

AspS9 I

AsuC2 I

AsuHP I

AsuNH I

BamH I

Bbv12 I

Bgl I

Bgl II

Bme18 I

Bpu14 I

Bsa29 I

Bsc4 I

Bse1 I

Bse3D I

Bse8 I

Bse21 I

Bse118 I

BseP I

BseX3 I

Bsp13 I

Bsp19 I

Bsp1720 I

BspA2 I

BssNA I

BssT1 I

Bst2B I

Bst2U I

Bst4C I

BstAC I

BstAP I

BstBA I

BstDE I

BstDS I

BstF5 I

BstH2 I

BstHP I

100

100

0-10

100

50-75

25-50

0-10

25-50

0-10

100

75-100

50-75

100

0-10

25-50

50-75

100

125-50

75-100

0-10

75-100

75-100

100

25-50

75-100

25-50

50-75

100

0-25

50-75

75-100

0-10

75-100

50-75

50-75

50-75

100

50-75

50-75

100

100

25-50

50-75

25-50

100

100

100

50-75

25-50

50-75

10-25

25-50

75-100

10-25

100

50-75

50-75

100

100

75-100

100

0-25

100

75-100

0-10

100

25-50

25-50

50-75

100

25-50

75-100

25-50

10-25

25-50

10-25

0-25

75-100

75-100

25-50

100

75-100

50-75

50-75

25-50

50-75

10-25

0-10

10-25

75-100

100

75-100

10-25

100

10-25

100

75-100

25-50

25-50

10-25

100

100

100

100

75-100

25-50

25-50

10-25

100

75-100

0-10

75-100

50-75

50-75

25-50

75-100

50-75

25-50

75-100

75-100

50-75

50-75

50-75

50-75

100

10-25

50-75

10-25

25-50

100

100

75-100

75-100

100

100

75-100

50-75

100

25-50

50-75

25-50

100

75-100

50-75

25-50

25-50

100

75-100

75-100

25-50

25-50

100

75-100

50-75

50-75

50-75

50-75

10-25

50-75

50-75

10-25

75-100

0-10

100

75-100

25-50

0-25

100

0-25

50-75

100

0-25

50-75

75-100

10-25

100

25-50

0-10

50-75

0-10

0-25

50-75

25-50

75-100

75-100

10-25

25-50

100

0-10

50-75

10-25

25-50

0-10

50-75

75-100

50-75

10-25

75-100

75-100

75-100

75-100

25-50

25-50

75-100

0-10

75-100

50-75

0-10

25-50

75-100

0-10

10-25

25-50

25-50

0-10

10-25

0-10

50-75

75-100

0-25

75-100

0-10

50-75

100

75-100

10-25

100

0-10

75-100

100

0-10

100

100

25-50

50-75

50-75

25-50

25-50

75-100

0-10

100

75-100

100

75-100

75-100

50-75

25-50

75-100

25-50

10-25

50-75

10-25

50-75

75-100

75-100

25-50

50-75

25-50

0-10

10-25

37

37

37

37

37

37

37

37

37

50

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

55

65

60

60

37

65

50

50

50

37

37

37

37

60

60

60

65

37

60

65

60

65

65

65

37

’

EnzymeOptimalBuffer B I O N R

OptimalTemp.

Activity in AccuCut buffers (%)

Restriction Enzymes

Enzymes

En

zym

es

112

N

N

R

N

O

I

O

I

O

I

EcoR I

B

N

B

N

I

I

I

R

O

B

R

N

I

N

O

O

R

N

N

B

N

R

B

N

I

N

O

N

I

I

I

I

R

O

N

R

I

R

O

N

N

I

R

O

N

I

B

I

O

R

B

R

I

N

N BstSE

Bst MS I

BstNS I

BstSF I

BstSN I

BstX2 I

Bsu6 I

BsuR I

CciN I

Dra I

DseD I

EcoR I

EcoR V

Ege I

Erh I

Fau I

FauND I

Fok I

FriO I

Fsp4H I

Hae III

Hind III

Hinf I

Hpa II

HspA I

Kpn I

Ksp22 I

Kzo9 I

Mlu I

Mly113 I

MroN I

MroX I

Msp I

MspR9 I

Nru I

NruG I

Ple19 I

Pme55 I

Psp124B I

PspE I

PspL I

PspN4 I

PspOM I

PspPP I

Pst I

Pvu I I

Rsa I

Sal I

Sbf I

SfaN I

Sfi I

Sfr274 I

Sfr303 I

Sma I

Smi I

Sph I

Sse9 I

Ssp I

Tru9 I

Tth111 I

Vha464 I

Vne I

Vsp I

Xba I

Xma I

Zsp2 I

N BstSE

10-25

10-25

50-75

10-25

10-25

10-25

50-75

75-100

75-100

50-75

100

100

50-75

100

0-10

50-75

25-50

0-10

75-100

50-75

100

75-100

25-50

25-50

25-50

50-75

50-75

25-50

10-25

0-10

100

75-100

50-75

100

10-25

25-50

10-25

0-10

50-75

10-25

25-50

0-10

10-25

25-50

25-50

50-75

25-50

0-10

75-100

25-50

50-75

10-25

0-10

75-100

75-100

50-75

50-75

100

10-25

10-25

25-50

100

50-75

0-10

25-50

10-25

50-75

75-100

50-75

50-75

25-50

100

50-75

100

75-100

100

50-75

25-50

75-100

10-25

50-75

100

100

100

25-50

50-75

0-10

75-100

50-75

100

75-100

25-50

50-75

10-25

50-75

10-25

25-50

75-100

75-100

10-25

75-100

100

25-50

75-100

50-75

100

100

100

100

25-50

25-50

75-100

0-10

100

0-10

25-50

75-100

75-100

100

25-50

50-75

75-100

100

50-75

100

75-100

25-50

25-50

75-100

100

25-50

0-10

75-100

50-75

25-50

50-75

100

50-75

100

50-75

100

75-100

75-100

25-50

75-100

25-50

25-50

75-100

50-75

75-100

50-75

100

25-50

75-100

50-75

50-75

25-50

100

100

10-25

25-50

50-75

50-75

75-100

50-75

10-25

25-50

75-100

50-75

100

50-75

75-100

10-25

10-25

25-50

25-50

100

50-75

10-25

50-75

25-50

100

75-100

50-75

0-10

25-50

100

75-100

50-75

25-50

50-75

100

25-50

10-25

75-100

50-75

50-75

0-10

100

100

75-100

100

75-100

50-75

75-100

25-50

75-100

75-100

50-75

0-10

100

10-25

100

50-75

50-75

75-100

50-75

75-100

10-25

25-50

100

50-75

100

50-75

50-75

0-10

100

100

50-75

100

50-75

0-10

100

75-100

100

75-100

100

75-100

10-25

75-100

50-75

10-25

25-50

100

0-10

75-100

10-25

75-100

100

100

0-10

25-50

25-50

100

75-100

75-100

75-100

50-75

10-25

0-10

75-100

75-100

100

0-10

10-25

10-25

100

0-10

0-10

10-25

25-50

75-100

25-50

25-50

75-100

50-75

10-25

25-50

0-10

10-25

25-50

10-25

100

25-50

0-10

100

10-25

25-50

25-50

50-75

50-75

100

10-25

10-25

50-75

50-75

100

75-100

10-25

25-50

10-25

10-25

25-50

25-50

10-25

0-10

10-25

100

25-50

0-10

100

0-10

100

25-50

50-75

10-25

0-10

100

75-100

50-75

25-50

25-50

10-25

25-50

100

50-75

100

0

25-50

10-25

50

37

60

65

60

37

37

37

37

37

37

37

37

37

55

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

37

50

50

37

25

37

37

55

37

65

65

37

37

37

37

37

37

55

Enzymes

En

zyme

s

113

Restriction Enzymes Isoschizomer Contents

Acc16 I

Acc65 I

Acc113 I

AccB1 I

AccB7 I

AccBS I

Acl I

AclN I

AclW I

Acs I

Afe I

Alu I

Ama87 I

Apa I

AsiA I

AspLE I

AspS9 I

AsuC2 I

AsuHP I

AsuNH I

BamH I

Bbv12 I

Bgl I

Bgl II

Bme18 I

Bpu14 I

Bsa29 I

Bsc4 I

Bse1 I

Bse3D I

Bse8 I

Bse21 I

Bse118 I

BseP I

BseX3 I

Bsp13 I

Bsp19 I

Bsp1720 I

BspA2 I

BssNA I

BssT1 I

Bst2B I

Bst2U I

BstAC I

BstAP I

BstBA I

BstDE I

BstDS I

BstF5 I

BstH2 I

BstHP I

BstMC I

BstNS I

BstSF I

BstSN I

BstX2 I

Bsu6 I

BsuR I

Aos I, Avi II, Fsp I, Mst I, Nsb I

Asp718 I, Sth I

Dpa I, Eco255 I, Sca I

Ban I, Hgic I, BbvB I, Eco64 I, MspB4 I

Acp II, Asp10H II, Esp1396 I, Van91 I, PflM I

BsrB I, Mbi I

Psp1406 I

Spe I

Bin I, Alw I, BspP I

Apo I, Fsi I, Xap I

Ait I, Aor51H I, Fun I, Eco47 III

Mlt I

Aqu I, Ava I, Bco I, BsoB I, Eco27K I, Eco88 I, NspII, NspSA I

Ppe I

Age I, PinA I, BshT I

Cfo I, FnuD III, Hha I

Asu I, Avc I, BspB II, Bsu54 I, Cfr13 I, Sau96 I

Aha I, Bcn I, Cau II, HgiS22 I, Nci I

Hph I

Nhe I, PstNH I

AccEB I, Ali I, Bna I, Bst I, NspSA IV, Sur I

Alw21 I, AspH I, Bsh45 I, HgiA I, BsiHKA I

Tsp8E I

Ncr I, NspMAC I, Pae2k I, Pae18k I

Afl I, Ava II, Bme216 I, Cau I, Eco47 I, HgiB I, HgiE I, Sin I

Acp I, Asp10H I, Asu II, Bsp119 I, BstB I, Csp45 I, Fsp II, Lsp I, Nsp V, Sfu I, Ssp1 I

Aag I, Ban III, Bci29 I, Bsc I, Bsp106 I, Bsu15 I, Cla I

BsiY I, Bsl I, BsaL I

BseN I, Bsr I, BsrS I, Bst11 I, Tsp1 I

BsrD I, BseM I

BsaB I, Bsh1365 I, BsiB I, BsrBR I, Mam I

Aoc I, Axy I, Bst29 I, Bst30 I, Bsu36 I, Cvn I, Eco81 I, Mst II,Sau I, SshA I

Bco118, BsrF I, BssA I, Cfr10 I

BssH II, Pau I

Aaa I, BstZ I, Eag I, EclX I, Eco52 I, Xma III

Acc III, BseA I, BsiM I, BspE I, BspM II, Bsu23 I, Kpn2 I, Mro I, Pta I

Nco I

Blp I, Bpu1102 I, Cel II, Esp I

Avr II, AvrB II, Bln I

BspM90 I, BstBS I, Bst1107 I, Sna I, Xca I

EcoT14 I, Eco130 I, Erh I, ErhB9 II, Sty I

Bsi I, BssSI

Aor I, Apy I, BseB I, Bse16 I, Bse17 I, Bse24 I, BspN I, Bst2 I, BstN I, BstO I, EcoR II,Fsp1604 I, Mva I,

Sth117 I, Zan I

Acy I, Aha I, Asu III, Bbi II, BsaH I, Hgi I, Hin1 I, Msp17 I, Pam II

ApaB I

BsaA I, MspY I

Dde I

Dsa I

BseG I

AccB2 I, Bsp143 II, Hae II

Hpa I

BsaO I, Bsh1285 I, BsiE I, Mcr I

Nsp I, NspH I

BdisS I, LlaB I, Sfc I, Sfe I

Eco105 I, Sna B I

BstY I, Mfl I, Tru201 I, Xho II

Bco5 I, Bco116 I, BseZ I, Ear I, Ksp632 I

Bim19 II, Bsh I, BspK I, BspR I, Bsp211 I, Dsa II, FnuD I, Hae III, Pla I, Sbv I, Sfa I

TGC GCA

G GTACC

AGT ACT

G G(C/T)(G/A)CC

CCANNNN NTGG

GAG CGG

AA CGTT

A CTAGT

GGATCNNNN

(G/A) AATT(C/T)

AGC GCT

AG CT

C (C/T)CG(G/A)G

GGGCC C

A CCGGT

GCG C

G GNCC

CC (C/G)GG

GGTGANNNNNNNN

G CTAGC

G GATCC

G(A/T)GC(A/T) C

GCCNNNN NGGC

A GATCT

G G(A/T)CC

TT CGAA

AT CGAT

CCNNNNN NNGG

ACTG G

GCAATGNN

GATNN NNATC

CC TNAGG

(G/A) CCGG(C/T)

G CGCGC

C GGCCG

T CCGGA

C CATGG

GC TNAGC

C CTAGG

GTA TAC

C C(A/T)(A/T)GG

C TCGTG

CC (A/T)GG

G(G/A) CG(C/T)C

GCANNNN NTGC

(C/T)AC GT(G/A)

C TNAG

C C(A/G)(C/T)GG

GGATGNN

(G/A)GCGC (C/T)

GTT AAC

CG(G/A)(C/T) CG

(G/A)CATG (C/T)

C T(G/A)(C/T)AG

TAC GTA

(G/A) GATC(C/T)

CTCTTCN

GG CC

BioneerEnzyme

Isoschizomer

Enzymes

En

zym

es

114

CciN I

Dra I

DseD I

EcoR I

EcoR V

Ege I

Erh I

Fau I

FauND I

Fok I

FriO I

Fsp4H I

Hae III

Hind III

Hinf I

Hpa II

HspA I

Kpn I

Ksp22 I

Kzo9 I

Mlu I

Mly113 I

MroN I

MroX I

Msp I

MspR9 I

Nru I

NruG I

Ple19 I

Pme55 I

Psp124B I

PspE I

PspL I

PspN4 I

PspOM I

PspPP I

Pst I

Pvu II

Rsa I

Sal I

Sbf I

SfaN I

Sfi I

Sfr274 I

Sfr303 I

Sma I

Smi I

Sph I

Sse9 I

Ssp I

Tru9 I

Tth111 I

Vha464 I

Vne I

Vsp I

Xba I

Xma I

Zsp2 I

Not I

Aha III

Drd I

Hal I, Kpn49k I, Rsr I, Sso I

Ceq I, Eco32 I

Ehe I, Eco78 I

BssT1 I, EcoT14 I, Eco130 I, ErhB9 II, Sty I

None

Nde I

None

Ban II, Bsu1854 I, Bsp519 I, Bvu I, Eco24 I, Eco215 I, HgiJ II, SacN I

BsoF I, Bsp6 I, Fbr I, Fnu4H I, Ita I, Uur960 I

Bim19 II, Bsh I, BspK I, Bsp211 I, BsuR I, Dsa II, FnuD I, Pla I, Sbv I, Sfa I

BstF I, EcoV III, Hsu I, Ssb I

CviB I, FnuA I, Hha II

Bco27 I, BsiS I, Bst40 I, Hap II, Msp I, Sth134 I

HinP1 I, Hin6 I, SciN I

None

AtuC I, Bco102 I, BspX II, Fba I, Pov I

AspMD I, BspA I, Bsp105 I, Bsp143 I, BtK II, Dpn II, Mbo I, Nde II, Nla II, Sau3A I

None

Mch I, Nar I, Nda I, Nun II, SseA I

Eco56 I, NgoA IV, NgoM I

Asp700 I, BbuA I, Xmn I

Hpa II

Bme1390 I, Msp67 I, ScrF I

Bsp68 I, MluB2 I, Sbo13 I, Spo I

Ahd I, AspE I, Eam1105 I, EclHK I, Uba1190 I, Uba1191 I

BspC I, ErhB9 I, Nb1 I, Ple19 I, Pvu I, Rsh I, Xor II

Aat I, Eco147 I, SseB I, Stu I

Sac I, Sst I

Acr II, AspA I, BstE II, BstP I, Eca I, EcoO65 I, Eco91 I, NspSA II

BpuB5 I, BsiW I, Pfl23 II, PpuA I, Sp1 I,Sun I

AspN I, BscB I, Nla IV

Bsp120 I

Pfl27 I, PpuM I, Psp5 II

Api I, Asp713 I, BspB I, Bsp63 I, Hal II, Sfl I

Bav I, Dma I, Pvu84 II

Afa I

HgiC III, HgiD II, Nop I, Xci I

Sse8387 I

None

Sdi I

Abr I, Blu I, Ccr I, Mav I, PaeR7 I, Pan I, Sla I, Xho I, Xpa I

Cfr42 I, Csc I, Gal I, Kpn19 I, Ksp I, Sac II, Sst II

CfrJ4 I, PaeB I, PspAL I

Swa I

Bbu I, Pae I

TspE I, Tsp509 I

unfound

Mse I, Tru1 I

Asp I, Ats I

Afl II, Bfr I, BspT I, Bst98 I, Esp4 I, MspC I

Aaq I, Alw44 I, ApaL I, Sno I

Ase I, Asn I, PshB I, Vsp I

None

Ahy I, Cfr9 I, EaeA I, PspA I, Xcy I, XmaC I

EcoT22 I, Mph1103 I, Nsi I, PinB I, Sep I

GC GGCCGC

TTT AAA

GACNNNN NNGTC

G AATTC

GAT ATC

GGC GCC

C C(A/T)(A/T)GG

CCCGCNNNN

CA TATG

GGATG(9)

G(G/A)GC(C/T) C

GC NGC

GG CC

A AGCTT

G ANTC

C CGG

G CGC

GGTAC C

T GATCA

GATC

A CGCGT

GG CGCC

G CCGGC

GAANN NNTTC

C CGG

CC NGG

TCG CGA

GACNNN NNGTC

CGAT CG

AGG CCT

GAGCT C

G GTNACC

C GTACG

GGN NCC

G GGCCC

(G /A) GG(A/T)CC(C/T)

CTGCA G

CAG CTG

GT AC

G TCGAC

CCTGCA GG

GCATCNNNNN

GGCCNNNN NGGCC

C TCGAG

CCGC GG

CCC GGG

ATTT AAAT

GCATG C

AATT

AAT ATT

T TAA

GACN NNGTC

C TTAAG

G TGCAC

AT TAAT

T CTAGA

C CCGGG

ATGCA T

Enzymes

En

zyme

s

115

AATT ACGT AGCT ATAT CATG CCGG CGCG CTAG GATC GCGC GGCC GTAC TATA TCGA TGCA TTAA

Enzymes

En

zym

es

116

unit Cat No.

200 E-1111

1,000 E-1112

Source: Alcaligenes faecalis T2774.

Reaction Condition: AccuCut buffer Violet: 33 mM Tris-acetate, 10 mM Mg-acetate, 66 mM K-acetate, 1 mM DTT, pH 7.9,Incubate at 37 .

Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA,1 mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20

Concentration: *10,000 units/ml.

Heat Inactivation: Yes. (65 for 20 minutes)

Isoschizomer: Ait I, Aor51H I, Fun I, Eco 47 III.

Neoschizomer: Unfound

Reactivity on methylated substrate DNA: Unidentified

5' AGCGCT 3'3' TCGCGA 5'

unit Cat No.

200 E-1121

1,000 E-1122

Source: Alcaligenes faecalis T2774.


Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20



Isoschizomer: Ait I, Aor51H I, Fun I, Eco 47 III.



5' AGCT...3'3' TCGA...5'

unit Cat No.

2,500 E-1141

12,500 E-1142

Source: Acetobacter pasteurianus.

Reaction Condition: AccuCut buffer Violet : 33 mM Tris-acetate, 10mM Mg-acetate, 66 mM K-acetate, 1 mM DTT, pH 7.9, Incubate at 37 .

Storage Buffer : 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1 mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .

Concentration: *20,000-100,000 units/ml.


Isoschizomer: Ppe I.

Neoschizomer: PspOM I (GGGCCC).

Reactivity on methylated substrate

DNA: Blocked by overlapping dcm methylation (Cm5CWGG),

GGGCCm5C.

5' GGGCCC 3'3' CCCGGG 5'

unit Cat No.

, 1,000 E-2141

5,000 E-2142

Source: Vibrio species 343.

Reaction Condition: AccuCut buffer Blue: 10 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT, pH 8.5, Incubate at 37 .

Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .

Concentration: *40000 units/ml.


Isoschizomer: Ase I, Asn I, PshB I, Vsp I.



5' ATTAAT 3'3' TAATTA 5'

Enzymes

En

zyme

s

117

unit Cat No.

5,000 E-1211

25,000 E-1212

Source: Bacillus amyloliquefaciens H.

Reaction Condition: AccuCut buffer Orange : 10 mM Tris-HCl,10 mM MgCl2, 50 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .

Storage Buffer: 10 mM KH2PO4, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .



Isoschizomer: AccEB I, Ali I, Bna I, Bst I, NspSA IV, Sur I



DNA: Not blocked by overlapping dam methylation Gm5ATC,

GGATCm5C, GGm6ATCC, GG m6ATCm5, GGATCm4C :

Blocked by GGA Tm4CC#, GGA tm5CC, GGAThm5Cm5C,

GGAhm5UCC.

5' GGATCC 3'3' CCTAGG 5'

unit Cat No.

1,000 E-1241

5,000 E-1242

Source: Bacillus globigii.

Reaction Condition: AccuCut buffer Red: 50 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .

Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1

mM 2- mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .


Heat Inactivation: No. (65 for 20 minutes)

Isoschizomer: Ncr I, NspMAC I, Pae2k I, Pae18k I.



DNA: Not blocked by GCm5CN5GGCb:

Blocked by Gm5CCN5GGC, GCCN5GGm5C GC m4CN5GGC.

5' AGATCT...3'3' TCTAGA 5'

unit Cat No.

200 E-2071

1,000 E-2072

Source: Streptomyces phaeochromogenes.

Reaction Condition: AccuCut buffer Orange : 10 mM Tris-HCl,10 mM MgCl2, 50 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .




Isoschizomer: Bbu I, Pae I.



DNA: Blocked by GCm6ATGC, Ghm5CATGhm5C. Not blocked by

GCATGm5C.

5' GCATGC 3'3' CGTACG 5'

unit Cat No.

2,000 E-1431

10,000 E-1432

Source: Bacillus stearothermophilus 2U.

Reaction Condition: AccuCut buffer Orange: 10 mM Tris-HCl,10 mM MgCl2, 50 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 60 .




Isoschizomer: Aor I, Apy I, BseB I, Bse16 I, Bse17 I, Bse24 I,BspN I, Bst2 I, BstN I, BstO I, EcoR II,Fsp1604 I,Mva I, Sth117 I, Zan I.



DNA: Not blocked by Cm5CWGG.

5' CC(A/T)GG 3'3' GG(T/A)CC 5'

Enzymes

En

zym

es

118

unit Cat No.

600 E-1271

3,000 E-1272

Source: Bacillus stearothermophilus 29.



Concentration: *15,000 units/m .


Isoschizomer: Aag I, Ban III, Bci 29 I, Bsc I,Bsp106 I, Bsu15 I, Cla I.



DNA: Blocked by Gm5ATC.

5' ATCGAT 3'3' TAGCTA 5'

unit Cat No.

400 E-1471

2,000 E-1472

Source: Bacillus stearothermophilus DE.


Storage Buffer: 10mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1 mM2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .



Isoschizomer: Dde I.



5' CTNAG 3'3' GANTC 5'

unit Cat No.

2,000 E-1601

10,000 E-1602

Source: Deinococcus radiophilus.


Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1mM EDTA, 50%glycerol, pH 7.5, 1 mM 2-mercaptoethanol, Store at -20



Isoschizomer: Aha III.



DNA: Blocked by TTTAm6AA

5' TTTAAA 3'3' AAATTT 5'

unit Cat No.

200 E-1781

1,000 E-1782

Source: Kurthia zopfil 9.





Isoschizomer: AspMD I, BspA I, Bsp105 I, Bsp143 I, BtK II,Dpn II, Mbo I, Nde II, Nla II, Sau 3A I

Neoschizomer: Dpn I (GATC).


DNA: Not blocked by Gm5ATC.

5' GATC 3'3' CTAG 5'

Enzymes

En

zyme

s

119

unit Cat No.

5,000 E-1621

25,000 E-1622

Source: Escherichia coli.

Reaction Condition: AccuCut buffer EcoR1: 100 mM Tris-HCl,10 mM MgCl2, 50 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .

Storage Buffer: 20 mM Tris-HCl, 300 mM KCl, 1 mM EDTA, 10mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20



Isoschizomer: Hal I, Kpn49k I, Rsr I, Sso I.



DNA: Blocked by Gm6ATTC, GA m6ATTC, GAAT Tm5C. Not

blocked by GAATThm5C, GAAhm5Uhm5UC.

unit Cat No.

2,000 E-1631

10,000 E-1632

Source : Escherichia coli.

Reaction Condition : AccuCut buffer Blue: 10 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT, pH 8.5, Incubate at 37 .




Isoschizomer: Ceq I, Eco 32 I.



DNA: Blocked by Gm6ATATC, GA Tm6ATC. Not blocked by GATA

Tm5C, GATAThm5C.

5' GAATTC 3'3' CTTAAG 5'

5' GATATC 3'3' CTATAG 5'

unit Cat No.

50 E-1661

250 E-1662

Source: Flavobacterium aquatili.

Reaction Condition: AccuCut buffer Green: 10 mM Tris-HCl,10 mM MgCl2, 1 mM DTT, pH 7.6, Incubate at 55 .




Isoschizomer: None .



5' CCCGCNNNN 3'3' GGGCGNNNNNN 5'a

unit Cat No.

3,000 E-1711

15,000 E-1712

Source: Haemophilus aegyptius.


Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1mM EDTA, 1 mM2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .


Heat Inactivation: No.(65 for 20 minutes)

Isoschizomer: Bim19 II, Bsh I, BspK I, Bsp211 I,BsuR I, Dsa II, FnuD I, Pla I, Sbv I,Sfa I



DNA: Blocked by GGm5CC. Not blocked by GGCm5C.

5' GGCC 3'3' CCGG 5'

Enzymes

En

zym

es

120

unit Cat No.

2,000 E-1731

10,000 E-1732

Source: Haemophilus influenzae.


Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1 mM2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .



Isoschizomer: CviB I, FnuA I, Hha II.



DNA: Blocked by Gm6ANTC, GANThm5C. Not blocked by GANT m5C.

5' GANTC 3'3' CTNAG 5'

unit Cat No.

1,000 E-1161

5,000 E-1162

Source: Arthrobacter species LE3860


Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercapto ethanol, 50% glycerol pH 7.5, Store at -20 .



Isoschizomer: Cfo I, Fnu D III, Hha I.

Neoschizomer: HinP I (GCGC), HspA I (GCGC).

Reactivity on methylated substrate DNA: unidentified.

5' GCGC...3'3' CGCG...5'

unit Cat No.

10,000 E-1721

50,000 E-1722

Source: Haemophilus influenzae Rd.

Reaction Condition : AccuCut buffer Blue: 10 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT, pH 8.5, Incubate at 37 .

Storage Buffer: 10 mM Tris-HCl, 300 mM Nacl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% 50% glycerol, pH 7.5,Store at -20 .



Isoschizomer: BstF I, EcoV III, Hsu I, Ssb I.



DNA: Blocked by AAGhm5CTT, AAGm5CTT, m6AAGCTT. Not

blocked by AAGChm5Uhm5U, A m6AGCTT.

5' AAGCTT...3'3' TTCGAA...5'

unit Cat No.

200 E-1831

1,000 E-1832

Source: Moraxella species.

Reaction Condition: AccuCut buffer Green : 10 mM Tris-HCl,10 mM MgCl2, 1 mM DTT, pH 7.6, Incubate at 37 .




Isoschizomer: Hpa II.



DNA: Blocked by m5CCGG, m5CCGG, m5Cm5CGG. Not blocked bym4CCGG, Cm4CGG, Cm5CGG.

5' CCGG 3'3' GGCC 5'

Enzymes

En

zyme

s

121

unit Cat No.

1,000 E-1741

5,000 E-1742

Source: Haemophilus parainfluenzae.





Isoschizomer: Bco27 I, Bsi S I, Bst40 I, Hap II, Msp I, Sth134 I.



DNA: Blocked by m4CCGG, m5CCGG, Cm4CGG, Cm5CGG,hm5Chm5CGG.

5' CCGG 3'

3' GGCC 5'

unit Cat No.

3,000 E-1761

15,000 E-1762

Source: Klebsiella pneumonia.





Isoschizomer: None.

Neoschizomer: Acc65 I, Asp718, Asp718 I (GGTACC)


DNA: Blocked by GG Tm6ACC, GGTAm4CC, GGTAm5Cm5C,

GGTACm4C. Not blocked by GGTAm5CC, GGTACm5C

5' GGTACC 3'3' CCATGG 5'

unit Cat No.

1000 E-1791

5,000 E-1792

Source: Micrococcus luteus.





Isoschizomer: None.



DNA: Blocked by Am5CGCGT. Not blocked by m6ACGCGT.

5' ACGCGT 3'3' TGCGCA 5'

unit Cat No.

200 E-2101

1,000 E-2102

Source: Thermus ruber 9.

Reaction Condition: AccuCut buffer Blue: 10 mM Tris-HCl, 10mM MgCl2, 100 mM KCl, 1 mM DTT, pH 8.5, Incubate at 65 .




Isoschizomer: Mse I, Tru1 I.



5' TTAA...3'3' AATT...5'

Enzymes

En

zym

es

122

unit Cat No.

200 E-1801

1,000 E-1802

Source: Micrococcus lylae 113.





Isoschizomer: Mch I, Nar I, Nda I, Nun II, SseA I.

Neoschizomer: Bbe I (GGCGCC), Ege I, Ehe I (GGCGCC), Kas I (GGCGCC)

Reactivity on methylated substrateDNA: Unidentified

5' GGCGCC 3'3' CCGCGG 5'

unit Cat No.

500 E-1671

2,500 E-1672

Source: Flavobacterium aquatili ND.

Reaction Condition: AccuCut buffer Violet : 33 mM Tris-acetate, 10 mM Mg-acetate, 66 mM K-acetate, 1 mM DTT, pH 7.9,Incubate at 37 .




Isoschizomer: Nde I.Neoschizomer: Unfound


5' CATATG 3'3' GTATAC 5'

unit Cat No.

500 E-1381

2,500 E-1382

Source: Bacillus species 19.

Reaction Condition: AccuCut buffer Blue: 10 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT, pH 8.5, Incubate at 37 .




Isoschizomer: Nco I.


Reactivity on methylated substrate DNA: unidentified

5' CCATGG 3'3' GGTACC 5'

unit Cat No.

600 E-1201

3,000 E-1202

Source: Actinobacillus suis NH.





Isoschizomer: Nhe I, Pst NH I.


Reactivity on methylated substrate DNA: unidentified

5' GCTAGC 3'3' CGATCG 5'

Enzymes

En

zyme

s

123

unit Cat No.

10,000 E-1951

50,000 E-1952

Source: Providencia stuartii.

Reaction Condition: AccuCut TM buffer Red: 50 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .




Isoschizomer: Api I, Asp713 I, BspB I, Bsp63 I, Hal II, Sfl I.


Reactivity on methylated substrateDNA: Blocked bym5CTGCAG/m5CTGm5CAG/CTGm5CAC/CTGCm6AG

unit Cat No.

1,500 E-1961

7,500 E-1962

Source: Proteus vulgaris 84.

Reaction Condition: AccuCut TM buffer Orange: 10 mM Tris-HCl,10 mM MgCl2, 50 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .




Isoschizomer: Bav I, Dma I, Pvu84 II.



DNA: Blocked by m5CAGCTG/CAGm5CTG/CAGm4C T G

5' CTGCAG 3'3' GACGTC 5'

5' CAGCTG 3'

3' GTCGAC 5'

unit Cat No.

200 E-1591

1,000 E-1592

Source: Curtobacterium citreus N.

Reaction Condition: AccuCut buffer Violet: 33 mM Tris-acetate,10 mM Mg-acetate, 66 mM K-acetate, 1 mM DTT, pH 7.9, Incubateat 37 .


Concentration: *5,000 units/ml. (Assayed Adenovirus-2 DNA)


Isoschizomer: Not I.



5' GCGGCCGC 3'3' CGCCGGCG 5'

unit Cat No.

3,000 E-2031

15,000 E-2032

Source: Streptomyces fradiae 274.





Isoschizomer: Abr I, Blu I, Ccr I, Mav I, PaeR7 I, Pan I, Sla I, Xho I, Xpa I.



5' CTCGAG 3'3' GAGCTC 5'

Enzymes

En

zym

es

124

unit Cat No. 5'. GCATCNNNNN 3'3'. CGTAGNNNNNNNNN 5'

Source: Streptococcus faecalis N.





Isoschizomer: None.



DNA: Not blocked by GCATm5C.

unit Cat No.

1,000 E-1971

5,000 E-1972

Source: Rhodopseudomonas sphaeroides.





Isoschizomer: Afa I.



DNA: Blocked by GTm6AC /GTA m4C. Not blocked by GTAm5C.

5' GTAC 3'3' CATG 5'

unit Cat No.

1,500 E-1891

7,500 E-1892

Source: Pseudomonas species 124B.





Isoschizomer: Sac I, Sst I.

Neoschizomer: Ecl136 II, EcoICR I (GAGCTC).


5' GAGCTC 3'3' CTCGAG 5'

unit Cat No.

1,000 E-1981

5,000 E-1982

Source: Streptomyces albus.

Reaction Condition: AccuCut buffer Red: 50 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT pH 7.6, Incubate at 37 .

Storage Buffer: 10mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1 mM2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20



Isoschizomer: HgiC III, HgiD II, Nop I, Xci I.



DNA: Blocked by G tm5CGAC, GTCGm6AC, Ghm5UCGAC. Not

blocked by GTCGAm5C.

5' GTCGAC 3'3' CAGCTG 5'

50 E-2001

250 E-2002

Enzymes

En

zyme

s

125

unit Cat No.

1,000 E-2051

5,000 E-2052

Source: Serratia marcescens.

Reaction Condition : AccuCut buffer Violet: 33 mM Tris-acetate, 10 mM Mg-acetate, 66 mM K-acetate, 1 mM DTT, pH 7.9,Incubate at 25 .


Concentration :*25,000 units/ml.


Isoschizomer: CfrJ4 I, PaeB I, PspAL I.

Neoschizomer: Cfr9 I, Xma I (CCCGGG)


DNA: Bolcked by m4CCCGGG, m5CCCGGG, Cm4CCGGG,

CCm4CGGG, CCm5CGGG. Not blocked by Cm5CCGGG.

5' CCCGGG 3'3' GGGCCC 5'

unit Cat No.

2,000 E-2151

10,000 E-2152

Source: Xanthomonas badrii.





Isoschizomer: None.



DNA: Blocked by TCTAGm6A, Tm5CTAGA, Thm5CTAGA.

5' TCTAGA 3'3' AGATCT 5'

unit Cat No.

50 E-2161

250 E-2162

Source: Xanthomonas malvacearum





Isoschizomer: Ahy I, Cfr9 I, EaeA I, PspA I, Xcy I, XmaC I.

Neoschizomer: Sma I


5 CCCGGG 33 GGGCCC 5

Documents

bioneer catalog 2006 - Bioneer Catalog 2006.pdf · Enzymes Enzymes 98 Fig 2. Extension time was limited to 60 sec and 1 ~ 10 kb of lambda DNA target gene was amplified. Bioneerss