A simple and robust method for detecting DNA alterations
Surveyor Mutation Detection Kits allow quick and easy detection of mutations and polymorphisms in DNA. The key component of the kits, Surveyor Nuclease, a member of the CEL family of mismatch-specific nucleases, recognizes and cleaves mismatches due to the presence of single nucleotide polymorphisms (SNPs) or small insertions or deletions.
For detecting CRISPR genome editing events, we recommend the Alt-R® Genome Editing Detection
Surveyor Mutation Detection Kits provide a simple and robust method for detecting mutations and polymorphisms in DNA from a variety of organisms, including bacteria, fungi, plants, and animals. The key component of the kits is Surveyor Nuclease, a member of the CEL family of mismatch-specific nucleases derived from celery. Surveyor Nuclease recognizes and cleaves mismatches due to the presence of single nucleotide polymorphisms (SNPs) or small insertions or deletions. Your PCR products undergoing analysis by Surveyor Nuclease should be prepared with a high fidelity thermostable DNA polymerase, such as Optimase® Polymerase (Transgenomic), to minimize the incorporation of errors that would result in background mismatches. For standard assays, T-Taq DNA Polymerase can also be used.
The Surveyor Mutation Detection Kit for Standard Gel Electrophoresis has been designed to cleave unlabeled DNA fragments at mismatched sites for subsequent analysis by agarose gel electrophoresis or polyacrylamide gel electrophoresis (PAGE). DNA fragments 200–4000 bp in length can be analyzed using manual agarose gel electrophoresis, while smaller fragments (<1000 bp) can be analyzed using manual polyacrylamide gel electrophoresis (PAGE). Reaction buffers and positive controls included in the kit allow you to set up and monitor the assays to obtain reproducible results.
Surveyor kits are shipped on ice and should be stored at –20˚C in a non–frost-free freezer. Enzyme quality is guaranteed for 6 months from receipt when stored as directed.
Figure 1. Accurate detection of mutations. Fragments generated by digestion of 160 ng of DNA with 1 μL of Surveyor Nuclease S, at 42°C for 20 min, were analyzed by electrophoresis on a 2.5% agarose gel containing ethidium bromide, using 75% of the digested DNA. The 584 bp amplicons were generated by PCR amplification with Optimase Polymerase (Transgenomic) from 1 wild-type reference and 3 mutant genes. Mutant 1 (#26): T>C—fragments = 242 and 342 bp; mutant 2 (#3): A>G and T>C—fragments = 135 and 449 bp, 220 and 364 bp; and mutant 3 (#13): single-base deletion, T>C, and C>A—fragments = 120 and 464 bp,170 and 414 bp, and 226 and 358 bp. A 100 bp ladder was run as a sizing standard.