cDNA normalization

Construction of normalized cDNA libraries: terms and prices

cat.# CS011

Starting material

Eukaryotic total RNA (3-5 μg) or eukaryotic poly(A)+ RNA (1-2 μg)

Service details (service levels may be adjusted for your particular needs)

PROCEDURE / CAT.#
CS011-2A
LEVEL 2A
CS011-2B
LEVEL 2B
CS011-2C
LEVEL 2C
CS011-3A
LEVEL 3A
CS011-3B
LEVEL 3B
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.

NOTES:
*Library cloning is performed into our standard vector analogous to pGEM-T vector using SfiIA – SfiIB restriction sites. Cloning using other sites or into a customer-provided vector currently can not be performed.
**To prepare an amplified cDNA library, the non-amplified library is arrayed in five 25x25 cm plates (approximately 100 000 colonies per plate); clones are eluted by LB/Amp medium; then the library is supplied with 17% glycerol and stored at -70°C. Preparation of the normalized amplified cDNA library containing more than 400 000 independent clones can be performed at the agreed price.
1. Synthesis of ds cDNA suitable for directional cloning
2. DSN-normalization of cDNA
3. Ligation of normalized cDNA into a vector*
4. PCR quality control of 22 randomly picked clones
5a. Preparation of non-amplified normalized cDNA library containing about 100 000 independent clones
5b. Preparation of amplified normalized cDNA library containing about 400 000 independent clones**
6. Purification of plasmid DNA from the 90 clones
7. Single run sequencing of 3'-ends of the 90 clones
Turnaround time, weeks
3-5
4-6
4-6
5-7
5-7
Price
€ 2 700
€ 3 500
€ 3 500
€ 4 500
€ 4 500

Deliverables

OUTPUT MATERIALS / CAT.#
CS011-2A
LEVEL 2A
CS011-2B
LEVEL 2B
CS011-2C
LEVEL 2C
CS011-3A
LEVEL 3A
CS011-3B
LEVEL 3B
1. Amplified non-normalized cDNA (1 μg)
2. Normalized cDNA (2 μg)
3. PCR primers for cDNA amplification
4. Service report
5. Normalized cDNA ligated into a vector
6. Non-amplified normalized cDNA library (about 100 000 independent clones)
7. Amplified normalized cDNA library (about 400 000 independent clones)
8. Purified plasmid DNA from the 90 clones and insert sequencing data of these 90 clones

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