突变位点序列确认

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 using nuclease based genome editing technologies, including CRISPR/Cas9, TALENs, and zinc finger nucleases, in a cell population. 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.