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
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.