[A method for the preparation of normalized cDNA libraries enriched with full-length sequences].
We developed a new method for the preparation of normalized cDNA libraries enriched with full-length sequences. It is based on the properties of the recently characterized duplex-specific nuclease from the hepatopancreas of the Kamchatka crab. The duplex-specific nuclease is thermostable, it effectively cleaves double-stranded DNA and is inactive toward single-stranded DNA (Shagin et al., Genome Res., 2002, vol. 12, pp. 1935-1942). Our method enables the normalization of cDNA samples enriched with full-length sequences without use of laborious and ineffective stages of physical separation. The efficiency of the method was demonstrated in model experiments using cDNA samples from several human tissues.
Zhulidov
P A
PA
Bogdanova
E A
EA
Shcheglov
A S
AS
Shagina
I A
IA
Vagner
L L
LL
Khazpekov
G L
GL
Kozhemiako
V V
VV
Luk'ianov
S A
SA
Shagin
D A
DA
D000818:Animals; D003386:Brachyura; D018076:DNA, Complementary / Q000737:chemistry*; D004277:DNA, Single-Stranded / Q000737:chemistry*; D004720:Endonucleases / Q000737:chemistry; D015723:Gene Library; D006801:Humans; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction
D018076:DNA, Complementary; D004277:DNA, Single-Stranded; D004720:Endonucleases