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CRISPR Integration Kit

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recombinant

expressed in E. coli

Quality Level

packaging

pkg of (6 vials per kit)

application(s)

CRISPR
genome editing

selection

kanamycin

shipped in

dry ice

storage temp.

−20°C

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CRISPRIS09CRISPRIS02CRISPRIS05
shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

storage temp.

−20°C

storage temp.

−70°C

storage temp.

−70°C

storage temp.

−70°C

Quality Level

200

Quality Level

-

Quality Level

-

Quality Level

-

recombinant

expressed in E. coli

recombinant

-

recombinant

-

recombinant

-

application(s)

CRISPR
genome editing

application(s)

CRISPR

application(s)

CRISPR

application(s)

CRISPR

General description

The CRISPR Integration Kit provides the essential genome editing reagents necessary to integrate a BstNI restriction site to the human KRAS locus. This kit is intended to serve as a tool to evaluate cell lines for ability to integrate new sequences at CRISPR-effected double strand breaks with the aid of short oligonucleotide donor. Integration efficiencies obtained with this kit enable rational design of genome editing experiments in a wide range of cell lines.

Application

CRISPR/Cas9 can be used to introduce, or “knock in”, new DNA sequences. Common modifications include the introduction of a SNP, small tag, loxP or larger cassette such as fluorescent protein. These modifications are made through the introduction of targeted DSB which enhance targeted integration significantly (Choulika et al., 1995). Targeted integration (gene knock in) occurs through homology directed repair (HDR)( (Bibikova et al., 2002). In order to enable gene editing by HDR, a DNA ‘donor′ or repair template containing the desired sequence must be delivered to the cell along with gRNA and Cas9.

Features and Benefits

  • The CRISPR Integration Assay provides a validated CRISPR that cleaves DNA with high efficiency along with a standardized oligo donor designed to survey homology dependent repair frequencies in a wide variety of cell lines
  • CRISPR-aided integration increases the targeted mutagenesis rate without the need for antibiotic selection
  • Integration efficiencies estimated with this kit enable rational design of genome editing experiments in a wide range of cell lines

Components

Each kit consists of:
one vial of gRNA plasmid - KRAS
one vial of Cas9 GFP plasmid
one vial of 120 base donor oligonucleotide
one vial of forward PCR primer
one vial of reverse PCR primer
one vial of genomic DNA control

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