cDNA normalization

Preparation of uncloned normalized cDNA for NG sequencing: terms and prices

cat.# CS010

Starting material

  • 3-5 μg eukaryotic total RNA
  • 1-2 μg eukaryotic poly(A)+ RNA
  • 3-5 μg prokaryotic total RNA

Service details

Different options are available for generating high-quality normalized cDNA preps suitable for further next generation ('NG') sequencing.

For all the options, cDNA synthesis is performed using modified template-switching (SMART) approach that allows introduction of known adapter sequences to the both ends of the first-strand cDNA. cDNA is further amplified by PCR and normalized using DSN-technology.

CAT.# Price
Service details
Starting material
Applications
Turnaround time is 2-4 weeks.
Prices include handling/shipping of service resulting materials (door-to-door delivery to one destination). Shipping of starting materials is paid by sender. We offer discounts for multiple orders.
CS010-1 € 1600 Full-length-enriched normalized cDNA is flanked on its 3'-end by adapter sequences comprising 1-4 (dT) stretches interspersed with dCs and dGs. This prevents problems with 454 sequencing as the cDNA does not contain long poly(A) stretches. The cDNA also comprises asymmetric SfiI sites and therefore suitable for directional cloning. Eukaryotic total or poly(A)+ RNA Equally well applicable for all NG sequencing platforms and for cloning projects.
Optimal for Roche 454 sequencing (Titanium chemistry)
CS010-1A € 1300 T-primer comprising 5-10 (dT) stretches interspersed with dC and dG is used for cDNA synthesis that helps to partially avoid 454 sequencing problems caused by presence of long homopolymer stretches in cDNA. Full-length-enriched normalized cDNA flanked by the same adapter sequences is suitable for NG sequencing (Roche 454, Illumina Solexa, ABI SOLiD) and for non-directional cloning. Eukaryotic total or poly(A)+ RNA Applicable for all NG sequencing platforms
CS010-1B € 1300 Full-length-enriched normalized cDNA is flanked by adapter sequences comprising asymmetric SfiI sites. SfiI enzyme digestion allows reducing size of the flanking fragments and generating sticky ends. cDNA can be used for NG sequencing and directional cloning. Eukaryotic total or poly(A)+ RNA Preferable for Solexa Illumina and ABI SOLiD sequencing.
Suitable for construction of full-length-enriched cDNA libraries for various needs
CS010-1C € 1300 T-primer with GsuI restriction site inserted is used for making first-strand cDNA; GsuI digestion reduces poly(A) tail in cDNA that help to get better 454 results. The option is tailored specifically for Roche 454 sequencing (both GS FLX Standard and Titanium Series as well as GS Junior System). Eukaryotic total or poly(A)+ RNA Preferable for Roche 454 sequencing

Deliverables

1. Amplified non-normalized cDNA (1 μg)

2. Amplified normalized cDNA (2 μg)

3. PCR primers for cDNA amplification

4. Leftover starting material (on request)

5. Service report

Extra options (please specify when ordering)

OPTION DESCRIPTION PRICE
Additional amplification of normalized/non-normalized cDNA (up to 30 μg) Amplification of cDNA by PCR with further phenol-chloroform extraction € 50 per 1 μg
Normalization efficiency analysis For human and mouse cDNA samples:
Virtual Northern blot is performed with ACTB and GAPDH genes-derived probes (high abundance in most tissues)
€ 500
For samples from other species:
PCR analysis: sequences of two abundant transcripts are required (provided by customer)
€ 200
Real-Time PCR analysis: sequences of two abundant transcripts are required (provided by customer) € 350
Virtual Northern blot: two cloned fragments corresponding to abundant transcripts are required (provided by customer) Request a quote

Important Notes

Turnaround time is estimated based on average turnaround time for this type of orders. Actual time may depend on the complexity of starting materials and/or deliverables.

In all the service procedures, outcomes are highly dependent on the nature and quality of starting materials. You will be advised on any concerns we may have related to samples quality after QC tests, and every effort will be made to obtain the best possible results.

Confidentiality statement

All data and materials derived from these services are the physical and intellectual property of the purchaser and will be held strictly confidential by EVROGEN.

Copyright 2002-2016 Evrogen. All rights reserved.
Evrogen JSC, 16/10 Miklukho-Maklaya str., Moscow, Russia, Tel +7(495)988-4084, Fax +7(495)988-4085, e-mail:evrogen@evrogen.com