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CRISPR/Cas9 Gene Editing Tools

CRISPR/Cas9 Gene Editing Tools - laboratoire.com Gene Editing Tool… · Gene editing for all researchers The CRISPR/Cas9 system has democratized genome modifi cation; targeted modifi

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Page 1: CRISPR/Cas9 Gene Editing Tools - laboratoire.com Gene Editing Tool… · Gene editing for all researchers The CRISPR/Cas9 system has democratized genome modifi cation; targeted modifi

CRISPR/Cas9Gene Editing Tools

Page 2: CRISPR/Cas9 Gene Editing Tools - laboratoire.com Gene Editing Tool… · Gene editing for all researchers The CRISPR/Cas9 system has democratized genome modifi cation; targeted modifi

2

Guide-it™ Products for Successful CRISPR/Cas9 Gene Editing

Gene editing for all researchersThe CRISPR/Cas9 system has democratized genome modifi cation; targeted modifi cation can now be achieved at virtually any genomic locus in virtually any cell type. This powerful method is allowing researchers to ask questions that were previously unaskable, leading to new insights into the basis of fundamental biological processes and to new innovative therapeutic strategies for treating disease—and that’s good science!

Best-in-class tools for your gene editing project—from start to fi nishGuide-it products further improve the usability of the CRISPR/Cas9 system by providing straightforward, streamlined methods at every step of your genome editing workfl ow.

-

Why choose Guide-it products?• Optimized methods

designed for speed and ease of use

• Complete kits that don’t require additional reagents

• Backed by our team of technical support scientists, ready to help at any point in your experiment Design sgRNAs

Transcribe and screensgRNAs in vitro

Deliver sgRNA and Cas9 to cells

Detect Cas9 protein and confi rm gene editing

Identify indels

Typical Gene Editing Workfl ow

Page 3: CRISPR/Cas9 Gene Editing Tools - laboratoire.com Gene Editing Tool… · Gene editing for all researchers The CRISPR/Cas9 system has democratized genome modifi cation; targeted modifi

www.takarabio.com/CRISPR3

What is CRISPR/Cas9 gene editing?CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 technology is based on an antiviral adaptive immune system in bacteria. The CRISPR/Cas9 system has been harnessed to create a simple, RNA-programmable method to mediate genome editing in mammalian cells. CRISPR/Cas9 editing is mediated by two components: a single-guide RNA (sgRNA) and Cas9 nuclease. The sgRNA directs the Cas9 nuclease to a specifi c genomic site where it introduces double-stranded DNA breaks. When this DNA damage is repaired by endog-enous cellular DNA repair mechanisms, mutant alleles can be introduced.

At the bench: Design the best sgRNAThe fi rst step in any CRISPR/Cas9 experiment is to choose a target sequence in the gene that you want to manipulate, and then design your guide RNA. In theory, selecting a target sequence is simple, but in practice, not all sgRNAs perform with equal effi ciency and specifi city. It is recommended that for any given target, multiple sgRNAs are designed and tested.

Learn more about best practices for choosing a target site at www.takarabio.com/sgRNA-design.

Design sgRNAs

sgRNA (single guide RNA)1 sgRNA + Cas9 protein2

Target-specificcrRNA sequence

tracrRNA

Your favourite gene

PAM sequence(5'-NGG-3')

Cas9

sgRNA

Target-specific cleavage3 Cellular repair4

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4

Test sgRNA performance before cell delivery

At the bench: Effective sgRNAs are identifi ed in vitro

Identifying effective sgRNAs. Panel A. Cleavage effi ciency of four different sgRNAs (1–4) targeting CXCR4. Negative control (NC) lacked sgRNA. sgRNA3 was predicted to be the least effective. Panel B. HeLa cells were cotransfected with plasmids encoding Cas9 and one of the four different sgRNAs. Effi ciency of CXCR4 gene disruption was assessed by a FITC-labeled antibody against CXCR4. In agreement with the Screening Kit results, sgRNA3 resulted in the lowest percentage of knockout cells.

Guide-it sgRNA In Vitro Transcription Kit: Produce high yields of sgRNAs• Fast, effi cient method for sgRNA production

• Transcribe 10-20 ug of sgRNA per in vitro reaction

• sgRNAs can be used for in vitro screening and/or delivery into cells via transfection/electroporation

Guide-it sgRNA In Vitro Screening Kit: Test sgRNA effi cacy• Highly optimized in vitro cleavage assay that allows

estimation of cleavage effi ciency

• Includes high-quality, recombinant Cas9 protein

200

Co

un

ts

160

120

80

40

0100 101

Knockout71.26%

102 103

FL1-H

sgRNA1

104

200

Co

un

ts

160

120

80

40

0100 101

wtwt wtwt

Knockout73.62%

102 103

FL1-H

sgRNA2

104

200

Co

un

ts

160

120

80

40

0100 101

Knockout11.50%

102 103

FL1-H

sgRNA3

104

200

Co

un

ts

160

120

80

40

0100 101

Knockout74.90%

102 103

FL1-H

sgRNA4

104

Transcribe and screensgRNA in vitro

Guide-it sgRNA Screening Kit in vitro Cas9 cleavage

assay

M NC 1 2 3 4 0% ~55% ~42% ~8% ~71%

A B

1 Design/purchase a 56- to 58-nt forward primer

2 Create template for in vitro transcription of sgRNA

3 Production of sgRNA by in vitro transcription and purification with the provided Guide-it IVT RNA Clean-Up Kit (Cat. No. 632638)

T7 promoter(17 nt + 4 nt)

Target-specificcrRNA (20 nt)

tracrRNA

Homology to scaffoldtemplate (15 nt)

56- to 58-ntforwardprimer Reverse primer

PCR

Scaffold template(tracrRNA)

Target-specificcrRNA

T7promoter

sgRNA

1 Use PCR to generate a target for cleavage

2 In vitro cleavage of target sequence by recombinant Cas9 and synthesized sgRNA

3 Separate cleavage products on an agarose gel

Uncleaved

Cleaved

PAM sequence(5'-NGG-3')

Cas9

sgRNA

Your target gene

PAMsequence

Page 5: CRISPR/Cas9 Gene Editing Tools - laboratoire.com Gene Editing Tool… · Gene editing for all researchers The CRISPR/Cas9 system has democratized genome modifi cation; targeted modifi

www.takarabio.com/CRISPR5

At the bench: Tight control ofCas9 expression

Lenti-XTM CRISPR/Cas9 Systems: Lentiviral delivery• Optimized systems for producing lentiviral particles to

deliver sgRNA/Cas9 expression cassettes

• Express Cas9 either constitutively or with doxycycline induction under control of the Tet-On® 3G promoter

– +200

Co

un

ts

160

120

80

40

0100 101

– Dox

102 103

M1M2

1.39%knockout

200

Co

un

ts

160

120

80

40

0100 101 102 103

M1M2

58.61%knockout

+ Dox

A Guide-it Mutation Detection Kit

B FACS

Doxycycline-induced genome editing in cell lines transduced with our Lenti-X Tet-On 3G CRISPR/Cas9 System. This clonal population of cells was transduced with both dox-inducible Cas9 and sgRNA pro- ducing lentiviral particles. As seen in panel A, cleavage of the target is seen after adding doxycycline (+), while there is no background genome editing before adding doxycycline (-). Panel B demonstrates functional editing of a surface membrane protein, with 59% knockout achieved after adding doxycycline.

Deliver sgRNA/Cas9 expression cassettes to any target cell

pGuide-it vector simultaneously expresses Cas9, an sgRNA, and a fl uores-cent reporter (ZsGreen1 or TdTomato).

Guide-it CRISPR/Cas9 Systems: Plasmid-based delivery • Complete system for the cloning and expression of

sgRNAs and Cas9 in mammalian cells

• Pre-linearized expression plasmids allow for simple, single-step cloning of sgRNAs

• Vectors include bright fl uorescent reporters, ZsGreen1 or tdTomato (2.5x and 6x brighter than EGFP, respectively) for identifi cation of transfected cells 

AAVpro® CRISPR/Cas9 Helper Free Systems: AAV-mediated delivery• Optimized systems for producing AAV2 particles to

deliver sgRNA/Cas9 expression cassettes. Packaging vectors for additional serotypes (AAV1, AAV5, AAV6) are available.

• Improved results in hard-to-transfect cells/suitable for in vivo use

• AAV delivery precludes genomic integration and persistent expression of Cas9, reducing potential off-target effects.

pAAV-Guide-it-Up Vector pAAV-Guide-it-Down Vector

Transfection+ pRC2-mi342 Vector+ pHelper Vector

HEK 293T Cells

AAV2-Up Particles AAV2-Down Particles

Cotransduction

Target cell

Cas9/sgRNAcomplex

Expression of full-length Cas9 & user-defined sgRNA

AAV Extraction Solution

HEK 293T Cells

Cas9

sgRNA

A two-vector system overcomes the size restrictions of the AAV genome. The SpCas9 gene is split between two vectors with a 1.6 kb region of homology that mediates recombination in target cells, producing full-length SpCas9.

Deliver sgRNA andCas9 to cells

• AAVpro CRISPR/Cas9 Helper Free System (with Streptococcus pyogenes Cas9): The SpCas9 gene is split between pAAV-Guide-it-Up and pAAV-Guide-it-Down plasmids with a 1.6-kb region of homology; the homologous region results in recombination in target cells, producing a full-length SpCas9 gene.

• AAVpro CRISPR/SaCas9 Helper Free System (with Staphylococcus aureus Cas9): The SaCas9 gene fi ts into one single plasmid that also contains the sgRNA expression cassette.

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At the bench: Functional knockout of the endogenous CD81 gene by sgRNA transfection

HT1080 cells stably expressing Cas9 (HT1080-Cas9) were transfected with 50 pmol of sgRNA targeting CD81, either once (1x) or twice (2x). Seven days later, cells were immunostained with a CD81 antibody (Ab) conjugated to an FITC fl uorophore and analyzed by fl ow cytometry. A control sample, comprised of HT1080-Cas9 cells, was analyzed by fl ow cytometry, either without (-Ab) or with (+Ab) the CD81 antibody. Both single and double transfection with sgRNA resulted in a shift in fl uorescence intensity, indicating successful CRISPR/Cas9-mediated knockout of CD81.

Xfect™ RNA Transfection Reagent: RNA delivery• Single reagent for transfection of cell lines and primary cells with mRNA and/or sgRNA• Very low cytotoxicity and high transfection effi ciency in primary cells• Simple, serum-compatible protocol

–Ab –sgRNA +Ab –sgRNA

99.89%

Co

un

ts

Co

un

ts

Co

un

ts

Co

un

ts

17.72%knockout

+Ab +sgRNA 2x+Ab +sgRNA 1x

19.43%knockout0.07%

Workfl ow of CRISPR/Cas9 Gesicle Production. The target-specifi c sgRNA is cloned into the provided plasmid, which is then mixed with the CRISPR/Cas9 Gesicle Packaging Mixes 1 and 2. Following a 10-minute incubation, the complete nanoparticle mix is applied to the Gesicle Producer 293T Cell Line in the presence of the provided A/C Heterodimerizer ligand. After 48–72 hours, gesicles containing active Cas9 protein complexed with your sgRNA can be collected and concentrated via centrifugation.

RNA and Protein delivery - minimize off-target effects

Guide-it CRISPR/Cas9 Gesicle Production System: Protein deliveryCRISPR/Cas9 gesicles are cell-derived nanovesicles containing active Cas9 protein complexed with a target-specifi c sgRNA. Gesicles are engineered with glycoproteins on their surface that mediate binding and fusion with the membranes of a wide range of target cells, allowing delivery of Cas9/sgRNA even to diffi cult-to-transfect and non-dividing cells. The CRISPR/Cas9 Gesicle Production System provides all components needed for cloning and expressing your target-specifi c guide RNA, and packaging reagents for producing CRISPR/Cas9 gesicles.

• Effi cient delivery of Cas9/sgRNA ribonucleoprotein (RNP) complexes to a broad range of cell types• Cas9 protein delivery eliminates genomic integration and reduces off-target effects• Tight control over dose and timing of delivery and editing

Add 10 μg ofplasmid encodingfor sgRNA (600 µl

final) to Mix 1

Vortex and addto Mix 2

Add A/C Heterodimerizer

Vortex andincubate 10 minutes

Apply to 293T cells

Incubate 48–72 hr

Collect media,centrifuge

Deliver sgRNA andCas9 to cells

Page 7: CRISPR/Cas9 Gene Editing Tools - laboratoire.com Gene Editing Tool… · Gene editing for all researchers The CRISPR/Cas9 system has democratized genome modifi cation; targeted modifi

www.takarabio.com/CRISPR7

Knock out of cell surface marker CD81 in induced pluripotent stem cells.  hiPSC-18 cells were electroporated with each vendor’s complete Cas9 ribonucleoprotein (RNP) system (the guide RNAs were produced either synthetically or with their in vitro transcription system), with all RNPs targeting the same sequence in CD81. Ten days after electroporation, cells were stained with a fl uorescent anti-CD81 antibody and then run on a FACS machine to detect CD81. The positive and negative staining controls show the FACS data from untreated cells with and without antibody staining. The Takara RNP system demonstrated very effi cient editing of the target compared to its competitors.

Guide-it Recombinant Cas9 (Electroporation-Ready): Protein delivery• Recombinant wild-type Streptococcus pyogenes Cas9 nuclease (SpCas9) with C-terminal nuclear-localization signal• High concentration (3 ug/ul) and low glycerol (10%)• Target hard-to-edit cells, including hematopoietic stem cells (HSCs) and human iPS cells (hiPSCs)• Electroporation of mammalian cells with Cas9-sgRNA ribonucleoprotein (RNP) complexes minimizes off-target effects

Deliver sgRNA andCas9 to cells

Start-To-Finish hiPSC Editing and Single-Cell-Cloning SystemsTraditionally, the establishment of a clonal population from edited hiPS cells grown and passaged as colonies is ineffi cient, challenging, and time consuming and often results in cell death or premature differentiation. The Cellartis® gene editing and single-cell cloning systems contain a defi ned culture system for effi cient single-cell cloning and expansion of edited hiPSC clones, based on Cellartis DEF-CS™.

• Cellartis iPSC Single-Cell Cloning DEF-CS Culture Media Kit: A complete system for effi cient expansion and scale-up manufacturing of hiPSCs in a feeder-free and defi ned environment. Superior single-cell survival Maintenance of pluripotency and stable karyotype

• Cellartis iPSC rCas9 Electroporation and Single-Cell Cloning System: A complete system that allows effi cient gene editing of hiPSCs via electroporation followed by single-cell cloning and expansion into 48-well plates. Recombinant Cas9, sgRNA In Vitro Transcription Kit, and Cellartis iPSC Single-Cell Cloning DEF-CS Culture Media Kit are included.

• Cellartis iPSC CRISPR/Cas9 Gesicle and Single-Cell Cloning System: A complete system that allows effi cient gene editing of hiPSCs via gesicle delivery of Cas9/sgRNA complexes followed by single-cell cloning and expansion into 48-well plates. Guide-it CRISPR/Cas9 Gesicle Production System and Cellartis iPSC Single-Cell Cloning DEF-CS Culture Media Kit are included.

At the bench: Effi cient CD81 knockout in hiPS cells cultured in DEF-CSHuman induced pluripotent stem cells (hiPSCs) growing in DEF-CS medium were electroporated with Cas9 in a ribonucleoprotein (RNP) complex with sgRNA specifi c to CD81. FACS analysis identifi ed that over 86% of the hiPSCs were CD81(-), indicating a high effi ciency in gene editing.

At the bench: Higher effi ciency compared to other vendor systems

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Deliver sgRNA andCas9 to cells

Guide-it Long ssDNA Production System: Produce ssDNA donor templates for knockinsThis kit uses an effi cient and simple in vitro protocol for the production of long single-stranded DNA (ssDNA) strands for use as repair template in knockin experiments. The kit provides a simple method for converting a dsDNA PCR product into ssDNA by selectively digesting either the sense or the antisense strand. The kit contains suffi cient reagents to create 50 ssDNA strands (25 pairs of sense and antisense strands).

• Create long ssDNA donor templates up to 5 kb• ssDNA can be co-electroporat ed with Cas9/sgRNA ribonucleoprotein (RNP) complexes for delivery into target cells• Benefi ts of using ssDNA as a template over dsDNA:

Drastically reduced tendency to randomly integrate into the genome, resulting in improved gene editing efficiency and lower background Low cytotoxic response to ssDNA template delivery No expression from non-integrated templates, making identification of correctly edited clones significantly easier

At the bench: ssDNA donor template production up to 5 kb

Production of ssDNA from a dsDNA template starting material. Long ssDNA was produced for tagging GADPH and Tyr genes with AcGFP1. The ssDNA has a smaller molecular weight than the corresponding dsDNA (Panel A). Panel B shows the successful production of ssDNA ranging from 2–5 kb for the CCR5 locus.

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www.takarabio.com/CRISPR9

A streamlined method for detecting mutationsGuide-it Mutation Detection Kit: Mismatch-based detection of indels• Complete system for confi rming the presence of indels

after genome editing

• Ultra-fast PCR-based method – direct amplifi cation from cells without genomic DNA extraction or sequencing

• Includes Guide-it Resolvase enzyme, a mismatch-specifi c nuclease that provides better specifi city than CEL1 nuclease

• Quick screen to determine the mutation frequency in your cell population, the entire protocol can be completed within 3-4 hrs

Guide-it Resolvase cleavesimperfectly matched DNA

Region of interest is amplified(mutation is marked in blue)

Amplify genomic DNA from cellsusing Terra™ PCR Direct Polymerase

1 Denature and reanneal2

Cleaved and uncleaved PCR fragments have different sizes

3 Separate cleaved and uncleavedPCR fragments using agarosegel electrophoresis

4Uncleaved

Cleavedfragment A

Cleavedfragment B

Uncleaved fragmentCleaved fragment ACleaved fragment B

At the bench: Improved mutation detection versus the CEL1 assay

HEK 293T cells were transfected with plasmids encoding Cas9 and a sgRNA targeting the AAVS1 locus. Mutations were easily discernible when using the Guide-it kit. In contrast, the CEL1 assay showed considerable smearing, making it diffi cult to determine cleavage effi ciency and reducing the ability to detect low levels of mutations.

Guide-it CEL1 nuclease assa y* M 1 2 3 4 5 6 M 1 2 3 4 5 6

bp

468 –

291– 177–

Uncleaved fragment

* Note: Smearing is due to non-specifi c cleavage

Cleavage productCleavage product

Sensitive detection of Cas9 expressionGuide-it Cas9 Monoclonal Antibody (Clone TG8C1)• Recognizes wild-type Cas9 from Streptococcus

pyogenes

• Suitable for western blotting and immunocytochemistry

• High sensitivity and specifi city: recognizes less than 1 ng Cas9 by Western Blot

At the bench: Wild-type and nickase mutant Cas9 are detected with high sensitivity

HEK 293 cells were transfected with plasmids encoding either wt or mutant Cas9 transcripts expressed from the CMV promoter; untransfected cells were used as a negative control. After 48 hours, the cells were lysed and Western blotting was performed. Blots were probed with primary anti-Cas9 polyclonal antibody (1:5,000 dilution). For detection, an HRP-conjugated goat anti-rabbit secondary antibody was used.

250 —

150 —

100 —

75 —

50 —

Unt

rans

fect

ed

wt C

as9

H840AD10A

H840A D10A

Cas9 mutants

H983A N863

Detect Cas9 protein and confi rmgene editing

Guide-it Cas9 Polyclonal Antibody• Recognizes wild-type Cas9, as well as nickase and

nuclease-defi cient versions

• Suitable for western blotting and immunocytochemistry

• High sensitivity and specifi city: recognizes as little as 0.15 ng of recombinant Cas9 by Western Blot

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Determine if one or both copies of your gene have indels

Characterize indels in four steps

Guide-it Genotype Confi rmation Kit: Simple detection of monallelic and biallelic mutations in single clones• Streamlined protocol that uses direct amplifi cation of

target genomic DNA from cells

• Includes highly purifi ed recombinant Cas9 nuclease for in vitro cleavage

Guide-it Indel Identifi cation Kit: Characterize indels at the sequence level• Complete kit contains all of the components needed to amplify, clone, and prepare modifi ed target sites for

DNA sequence analysis

• Ultra-fast protocol includes PCR amplifi cation directly from cells, and the In-Fusion® Cloning system for ligation-free cloning in 15 minutes

• Can be used to characterize the variety of indels in cell populations or to analyze single clones

INDELS

Cas9/sgRNAcleaves

both copies

wt MonoallelicMutation

BiallelicMutation

Cas9/sgRNAcleaves only

wt copy

No Cas9/sgRNA

cleavage

1 largefragment

1 large fragmentand

2 small fragments

2 small fragments

The Guide-it Genotype Confi rmation Kit includes a Cas9/sgRNA-mediated in vitro cleavage reaction for genotype determination after CRISPR/Cas9 gene editing. The same sgRNA that was used for gene editing must be used in the cleavage assay.

HEK 293 cells were treated with Cas9 and an sgRNA targeting the C4BPB gene to generate 15 single-cell clones. Panel A. The Guide-it Genotype Confi rmation Kit was used to determine the genotype, with wt, monoallelic (M), and biallelic (B) control reactions included in the analysis. Panel B. Sequencing results are shown for select clones from Panel A. Lowercase letters represent the wt sequence. As predicted, clone 2 is monoallelic and clones 7 and 9 are biallelic.

Identify indels

Controls

Clone #2

7

9

Individual clones after C4BPB editing

WT M B WT 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Target sitePAM

Target site

PAM

Target sitePAM

Target sitePAM11

A

B

At the bench: Accurate genotype determination in HEK 293 cells

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www.takarabio.com/CRISPR11

P R O D U C T SCat. # Product Size

Transcribe and screen sgRNAs in vitro

632636 Guide-it™ Complete sgRNA Screening System (includes Cat. # 632635, 632638, and 632639) 50 Rxns

632635 Guide-it™ sgRNA In Vitro Transcription Kit 50 Rxns

632638 Guide-it™ IVT RNA Clean-Up Kit 50 Rxns

632639 Guide-it™ sgRNA Screening Kit 50 Rxns

Deliver sgRNA and Cas9 into cells

632601 Guide-it™ CRISPR/Cas9 System (Green) 1 System

632602 Guide-it™ CRISPR/Cas9 System (Red) 1 System

632608 AAVpro® CRISPR/Cas9 Helper Free System (AAV2) 1 System

632609 AAVpro® CRISPR/Cas9 Vector System 1 System

632619 AAVpro® CRISPR/SaCas9 Helper Free System (AAV2) 1 System

632618 AAVpro® CRISPR/SaCas9 Vector System 1 System

632629 Lenti-X™ CRISPR/Cas9 System Each

632630 pLVX-hyg-sgRNA1 Vector System 10 Rxns

632631 pLVX-puro-Cas9 Vector 20 uL

632633 Lenti-X™ Tet-On® 3G CRISPR/Cas9 System 1 System

631450 Xfect™ RNA Transfection Reagent 1.2 mL

632613 Guide-it™ CRISPR/Cas9 Gesicle Production System (includes Cat. # 632612 and 632616) 1 System

632617 Gesicle Producer 293T Cell Line 1 mL

632612 pGuide-it-sgRNA1 Vector System 1 System

632616 Guide-it™ CRISPR/Cas9 Gesicle Packaging Set 10 Rxns

632642 Cellartis® iPSC CRISPR/Cas9 Gesicle and Single-Cell Cloning System 1 System

632640 Guide-it™ Recombinant Cas9 (Electroporation-Ready) 3 x 100 ug

632641 Guide-it™ Recombinant Cas9 (Electroporation-Ready) 100 ug

632643 Cellartis® iPSC rCas9 Electroporation and Single-Cell Cloning System 1 System

Y30021 Cellartis® iPSC Single-Cell Cloning DEF-CS™ Culture Media Kit 1 Kit

632644 Guide-it™ Long ssDNA Production System 25 Rxns

632645 Guide-it™ Long ssDNA Strandase Kit 25 Rxns

Confi rm Cas9 expression

632606 Guide-it™ Cas9 Polyclonal Antibody 3 x 100 uL

632607 Guide-it™ Cas9 Polyclonal Antibody 100 uL

632627 Guide-it™ Cas9 Monoclonal Antibody (Clone TG8C1) 3 x 100 ug

632628 Guide-it™ Cas9 Monoclonal Antibody (Clone TG8C1) 100 ug

Determine genome editing effi ciency in target cell population

631443 Guide-it™ Mutation Detection Kit 100 Rxns

631448 Guide-it™ Mutation Detection Kit 25 Rxns

Identify Indels

632611 Guide-it™ Genotype Confi rmation Kit 100 Rxns

631444 Guide-it™ Indel Identifi cation Kit 10 Rxns

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Gene Editing Resources

www.takarabio.com/CRISPR

Visit our website to explore our evolving portfolio of products, learning resources, and technical information for CRISPR/Cas9 gene editing.

• New products• Selection guides• Technical notes• Videos• Protocols• Learning resources

Takara Bio [email protected][email protected][email protected] • +33 139 046 880For Research Use Only. Not for use in diagnostic or therapeutic procedures. Not for resale. Takara and the Takara logo are trademarks of TAKARA HOLDINGS, Kyoto, Japan. AAVpro is a trademark of Takara Bio, Inc. Clontech, the Clontech logo, Guide-it, In-Fusion, that’s GOOD science!, Terra, and Xfect are trademarks of Takara Bio USA, Inc. All other marks are the property of their respective owners. Certain trademarks may not be registered in all jurisdictions.©2017 Takara Bio Europe

ZZGIBR0817www.takarabio.com