Amplification of cDNA ends based on template-switching effect and step-out PCR.
A new method for amplifying cDNA ends is described which requires only first-strand cDNA synthesis and a single PCR to generate a correct product with very low or no background. The method can be successfully applied to total RNA as well as poly A+ RNA. The same first-strand cDNA can be used to amplify flanking sequences of any cDNA species present in the sample.
Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, Moscow 117871, Russia.
M
M
Matz
D
D
Shagin
E
E
Bogdanova
O
O
Britanova
S
S
Lukyanov
L
L
Diatchenko
A
A
Chenchik
D018076:DNA, Complementary / Q000235:genetics*; D008957:Models, Genetic; D016133:Polymerase Chain Reaction / Q000379:methods*
D018076:DNA, Complementary