Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2011;729:85-98.
doi: 10.1007/978-1-61779-065-2_6.

Normalization of Full-Length-Enriched cDNA

Affiliations

Normalization of Full-Length-Enriched cDNA

Ekaterina A Bogdanova et al. Methods Mol Biol. .

Abstract

A well-recognized obstacle to efficient high-throughput analysis of cDNA libraries is the differential abundance of various transcripts in any particular cell type. Decreasing the prevalence of clones representing abundant transcripts before sequencing, using cDNA normalization, may significantly increase the efficacy of random sequencing and is essential for rare gene discovery. Duplex-specific nuclease (DSN) normalization allows the generation of normalized full-length-enriched cDNA libraries to permit a high gene discovery rate. The method is based on the unique properties of DSN from the Kamchatka crab and involves denaturation-reassociation of cDNA, degradation of the ds-fraction formed by abundant transcripts by DSN, and PCR amplification of the remaining ss-DNA fraction. The method has been evaluated in various plant and animal models.

Similar articles

See all similar articles

Cited by 6 articles

See all "Cited by" articles

Publication types

LinkOut - more resources

Feedback