cDNA normalization service

cat.#CS010, CS011

- Significant increase of the transcriptome sequencing efficacy
- Equalization of cDNA population prior library cloning
- Normalized cDNA is suitable for 454 sequencing
- Order processing is set up and monitored by the inventors of the technology

A great range in representation of different mRNAs in cells and tissues makes it extremely difficult to analyze rare message in cDNA libraries. Therefore, for rare transcript searching and analysis, creation of equalized (normalized) cDNA is required.

Our proprietary Duplex-Specific nualease (DSN)-based normalization is a high efficient and well-proved approach to equalize transcript concentration in a cDNA population enriched with full-length sequences. The flexibility of the normalization procedure allows simple modifications for various purposes.

Both total or poly(A)+ RNA is suitable for cDNA synthesis. Full-length-enriched cDNA is usually produced using SMART technology. The method combines cDNA synthesis and amplification and results in representative cDNA population enriched with full-length sequences even from small amounts of starting materials. After cDNA synthesis, the double stranded cDNA is normalized using DSN-normalization method.

Please see Terms and Prices for service variant you desire:

cDNA normaliaztion & construction of normalized full-length-enriched cDNA library: Terms & Prices	cDNA normaliaztion 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