NOVEL METHOD FOR DETECTION OF SMALL AMOUNTS OF RNA BASED ON THE SEMI-NESTED POLYMERASE CHAIN-REACTION

Citation
S. Sanders et Ga. Thomas, NOVEL METHOD FOR DETECTION OF SMALL AMOUNTS OF RNA BASED ON THE SEMI-NESTED POLYMERASE CHAIN-REACTION, Journal of clinical pathology-Molecular pathology, 50(2), 1997, pp. 105-107
Citations number
6
Categorie Soggetti
Pathology
Volume
50
Issue
2
Year of publication
1997
Pages
105 - 107
Database
ISI
SICI code
Abstract
A technique for the amplification of very small quantities of cDNA bas ed on a semi-nested polymerase chain reaction system has been develope d. The technique uses an additional blocking oligonucleotide ''primer' ' in the second round nest which is complementary to one of the first round primers. This prevents amplification of first round product or o f any other unwanted products dependent on that primer, but allows amp lification of second round product, using first round product as the t emplate. The second round primers are added through the oil layer, eli minating the possibility of introducing first round product aerosol in to the atmosphere, as would be the case in a fully nested system. Cont amination is therefore minimised, while amplification of the desired p roduct is maximised.