What is the procedure of the LO test?
The LO tests is “Yes” or “No” test, i.e.: The test is performed using 5′ 32P-labeled synthetic 17-mer single and double-stranded oligonucleotides as substrates which don’t have recognition sites for the restriction enzyme tested. 4 hour incubation is carried out with respective substrates and 10 units of the restriction enzyme in its reaction buffer. Reaction products are separated on a polyacrylamide gel and results are analyzed by autoradiography or storage phosphor imaging. The restriction enzyme tested is considered to be PureExtreme® when no traces of oligonucleotides’ degradation are visible. The LO test using single-stranded and double-stranded synthetic oligonucleotides and restriction enzymes from different vendors.
