Takara Guide-it Indel Identification Kit

The Guide-it Indel Identification Kit provides a complete workflow for identifying the variety of insertions and deletions (indels) created in a cell population using nuclease-based genome editing technologies, including CRISPR/Cas9, TALENs, and zinc finger nucleases. The kit contains all of the components needed to amplify, clone, and prepare modified target sites for DNA sequence analysis. The indel identification protocol in this kit has been optimized to provide quick and reliable results.

The Guide-it Indel Identification Kit provides a complete workflow for identifying the variety of insertions and deletions (indels) created in a cell population using nuclease-based genome editing technologies, including CRISPR/Cas9, TALENs, and zinc finger nucleases. The kit contains all of the components needed to amplify, clone, and prepare modified target sites for DNA sequence analysis. The indel identification protocol in this kit has been optimized to provide quick and reliable results. First, Terra PCR Direct polymerase is used to amplify a genomic DNA fragment containing the target site directly from crude cell lysates. The resulting amplified fragments are cloned directly, without restriction digestion, into a pre-linearized pUC19 vector using the In-Fusion Cloning system. After transformation into Stellar Competent Cells, colony PCR is used to amplify the target region from the plasmid. DNA sequence analysis of the PCR products is then used to identify indels generated at the targeted genomic site.

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Overview

• Streamlined method for characterizing the variety of indels introduced by gene editing technologies
• Ultrafast protocol includes Terra PCR Direct Polymerase for PCR amplification directly from cells and the In-Fusion Cloning system for ligation-free cloning in 15 minutes
• Ideal for identifying indels caused by Cas9/CRISPR, TALENs, or zinc-finger nuclease-based gene editing in a cell population
• Complete kit contains all of the components needed to amplify, clone, and prepare modified target sites for DNA sequence analysis

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Applications

• Detecting insertions and deletions introduced in mammalian genomic DNA
• Characterizing the variety of indels that are present in a cell population after nuclease-based gene editing

Components

• Guide-it Indel Identification Components

  • 110 µl Terra PCR Direct Polymerase Mix
  • 3 x 1 ml 2X Terra PCR Direct Buffer (with Mg2+, dNTP)
  • 400 µl Extraction Buffer 1
  • 40 µl Extraction Buffer 2
  • 10 µl pUC19 Cloning Vector, linearized (50 ng/µl)
  • 200 µl Colony PCR Forward Primer (15 µM)
  • 200 µl Colony PCR Reverse Primer (15 µM)
  • 6 x 1 ml PCR-Grade Water

• In-Fusion HD Cloning Kit

• NucleoSpin Gel and PCR Clean-up kit

• Stellar Competent Cells

Additional product information

Please see the product`s Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.