cDNA normalization

cDNA normalization service

cat.# CS010, CS011

- Normalized cDNA suitable for further sequencing by next generation technologies (Roche 454, Illumina Solexa, ABI SOLiD)
- Significant increase in transcriptome sequencing efficiency; especially helpful for high-throughput applications
- Equalization of cDNA population prior to library cloning

Substantial difference in mRNA representation in cells/tissues complicates sequence analysis of primary cDNA libraries, especially for gene discovery projects. Equalization of cDNA population overcomes prevalence of highly abundant transcripts and essentially facilitates discovery of rare genes.

Evrogen duplex-specific nuclease (DSN)-based normalization technology is a highly efficient and well proved approach to equalize transcript abundance in cDNA populations enriched with full-length sequences. High flexibility of the normalization procedure allows easy modifications for various purposes.

Both eukaryotic and prokaryotic RNA can be used as starting material for normalization procedure. Full-length-enriched cDNA is normally produced using modified template-switching (SMART) approach. The method combines cDNA synthesis and amplification and allows generating representative cDNA populations enriched with full-length sequences even from small amounts of starting material. After cDNA synthesis and amplification, double stranded cDNA is normalized using DSN-normalization method.

Uncloned normalized cDNA for Next Generation Sequencing
Eukaryotic total or poly(A)+ RNA as well as prokaryotic total RNA can be used as a starting material for generation of normalized cDNA samples.

Normalized cDNA libraries
Normalized full-length-enriched cDNA is directionally cloned into a plasmid vector from our collection (or into an appropriate customer-supplied plasmid vector) and transformed into E. coli. The service comes in various levels to meet your project needs.

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