cDNA normalization service

cat.# CS010, CS011

- Significant increase in transcriptome sequencing efficiency; especially helpful for high-throughput applications
- Equalization of cDNA population prior to library cloning
- Normalized cDNA is suitable for 454 and Solexa sequencing as well as SOLiD targeted re-sequencing
- Order processing is monitored by inventors of the technology

Substantial difference in mRNA representation in cells/tissues makes sequence analysis of primary cDNA libraries inefficient, especially for gene discovery projects. Creation of equalized (normalized) cDNA libraries greatly helps to discover rare transcripts.

Evrogen Duplex-Specific nuclease (DSN)-based normalization technology is a highly efficient and well proved approach to equalizing transcript abundance in cDNA populations enriched with full-length sequences. The flexibility of the normalization procedure allows simple modifications for various purposes.

Both total RNA and poly(A)+ RNA can be used as starting material for cDNA normalization procedure. Full-length-enriched cDNA is normally produced using modified SMART approach. The method combines cDNA synthesis and amplification and allows to generate representative cDNA populations enriched with full-length sequences even from small amounts of starting material. After cDNA synthesis/amplification step, double stranded cDNA is normalized using DSN-normalization method.

Please see Terms and Prices for service variant you desire:

cDNA normalization & construction of normalized full-length-enriched cDNA library: Terms & Prices	cDNA normalization for further 454 sequencing: Terms & Prices
cDNA is normalized, size fractionated, cloned into a plasmid vector from our collection (or into an appropriate customer-supplied plasmid vector), and transformed into E. coli.	Normalization results in a sharp decrease in the representation of the most abundant transcripts in cDNA samples, consequently enhancing the gene discovery rate of 454 sequencing. Preparing normalized cDNA over 2,5 μg is available!

	Normalization significantly increases gene discovery rate of cDNA library. Transcript distribution in standard and normalized cDNA libraries from Aplysia neurons: blue – unique sequences; green – non-unique sequences, yellow – all sequences.

Typical cDNA normalization result. (A) Agarose/EtBr gel-electrophoresis of non-normalized (1) and normalized (2) human cDNA samples; (B, C) concentration of abundant transcripts in these samples revealed by Virtual Northern blot. ACTB - β-actin; UBC - ubiquitin C; M, 1-kb DNA size markers (SibEnzyme); embr. - embryonic.

Depending on your particular needs we also offer the following custom cDNA libraries:

• Standard full-length-enriched cDNA libraries
• Depleted full-length-enriched cDNA libraries
• Subtracted cDNA libraries