Results obtained using Standard Gel Electrophoresis methods

SURVEYOR Nuclease cleavage products analyzed using standard gel electrophoresis. SURVEYOR Nuclease digestion products were derived from Control C homoduplex (lanes 1, 2, and 3) and Control G/C homoduplex/heteroduplex mixtures (lanes 4, 5, and 6), and analyzed by electrophoresis on a 2% TransOneK Agarose gel run in 1X TBE. The Control G/C homoduplex/heteroduplex mixture was formed by combining and hybridizing equal amounts of Control G and Control C homoduplex PCR products; it contains C/C and G/G mismatched heteroduplexes as well as homoduplexes. Lane M shows a 1 kb Plus Ladder (Invitrogen, Inc., Carlsbad, CA). The DNA substrates were prepared by PCR amplification using Optimase^® Polymerase. Various amounts of DNA (lanes 1 and 4, 280 ng in 4 µl; lanes 2 and 5, 560 ng in 8 µl; lanes 3 and 6, 840 ng in 12 µl) were digested directly in the PCR buffer after adding 1 µl of SURVEYOR Nuclease S and incubating the reaction for 20 min at 42°C.

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• Detection of all types of mismatches
• Detection of 1, 2, 3, 6, 9 and 12 bp insertions
• Detection 1, 2 or 3 mutations in a single heteroduplexed template
• Detection of mutations in a 2.95 kb template
• Detection of mutations in pools of PCR products
• Advantages of using a high fidelity thermostable DNA polymerase such as Optimase^® Polymerase
• A Practical Example - Detection of SNPs known to distinguish two strains of mice