Muts QC
Protocol
[edit]1. Anneal 4ul each of PCR product (from 30 ng/ul purified PCR product). PCR products are identical except for a 2 bp deletion:
9ul volume
Annealing buffer: 10mM Tris pH 8; 50 mM NaCl
Annealing protocol: 95C for 5 mins, 95 to 85 C at -2 C /second; 85 to 25 C at -0.1 C/second
2. Add 1 ul of 10x rxn buffer (1 M KCl; 500 mM Tris-HCl pH 8; 200 mM MgCl2; 1mM EDTA; 10 mM DTT; 20% glycerol)
3. Add 0.5 ul of Taq MutS (from stock original supply)
4. Incubate at 65 C for 30 mins.
5. Run on PAGE gel.
Lanes:
1 No MutS 2 Competing company MutS 3 Excellgen Catalog item MutS-MBP tag 4 Excellgen MutS lot P5582107 5 Excellgen MutS lot P7RW1.73 6 Excellgen MutS lot P7R2.4