SURVEYOR^® Check-It Kit for Clone Sequence Validation

The SURVEYOR Check-It Kit for Clone Sequence Validation has been specifically designed to analyze cloned DNA to detect mutations and errors which may have been introduced into insert sequences during the PCR amplification and cloning processes, or to verify the presence of desired mutations deliberately introduced by site-directed mutagenesis. The kits include SURVEYOR Nuclease, a mismatch-specific plant endonuclease that cleaves DNA heteroduplexes at all possible mismatch sites. The DNAs of prospective clones are PCR amplified directly from colonies. The resulting PCR products are then combined with similar reference fragments derived from reference cDNA or genomic DNA, or from the original reference clones, and heated and cooled to promote the formation of heteroduplexes if the sequences are different. After annealing to reference DNA and treatment of the duplexes with SURVEYOR Nuclease, the cleavage fragments are separated by standard agarose gel electrophoresis and visualized by staining with ethidium bromide or SYBR^® Gold.