MOLECULAR BIOLOGY SERVICES:

    cDNA subtraction

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Prokaryotic cDNA subtraction and library construction service: terms and prices

cat.# CS024

Starting material

3-5 μg prokaryotic total RNA

Service details

PROCEDURE / CAT.#
LEVEL 1
CS024-1
LEVEL 2
CS024-2
LEVEL 3
CS024-3
LEVEL 4
CS024-4
LEVEL 5
CS024-5
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.
1. ds cDNA preparation using random primers
2. Subtractive hybridization in both directions
3. Ligation of subtracted cDNAs into an appropriate vector
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4. Transformation of the ligate from step 3 into E. coli
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5. Plating of the two subtracted libraries in 96-well plates (X plates for one direction + Y plates for the other direction)
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1 + 1
5 + 1
5 + 5
6. Differential screening of the 96-well plates with two probes to determine the percentage of differentially expressed clones in each subtracted library (probes: forward subtracted cDNA, reverse subtracted cDNA)
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7. Purification of plasmid DNA from the differential clones – up to 100 plasmids per each library (up to 200 plasmids in total for each SSH / 2 libraries)
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8. Virtual Northern hybridization of 5 clones from each library to confirm differential expression
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9. Sequencing of confirmed differential clones (up to 10)
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Turnaround time, weeks
3-5
4-6
5-7
5-11
5-11
Price
Request a quote
Request a quote
Request a quote
Request a quote
Request a quote

Deliverables

OUTPUT MATERIALS / CAT.#
LEVEL 1
CS024-1
LEVEL 2
CS024-2
LEVEL 3
CS024-3
LEVEL 4
CS024-4
LEVEL 5
CS024-5
1. Amplified tester and driver cDNAs (0.5 μg)
2. Subtracted cDNA preps (0.5 μg)
3. Primers for amplification of subtracted cDNA and SSH products
4. Leftover starting material (on request)
5. Service report
6. Transformation-ready subtracted cDNA ligated into an appropriate vector
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7. 96-well plates with clones used for differential screening
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8. Purified plasmid DNA from differential clones selected (1.5 μg each)
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9. Differential screening and Virtual Northern blot data
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10. Subtracted cDNA libraries
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Extra options (please specify when ordering)

OPTION PRICE
Mirror orientation selection (MOS) € 1000
Addition amplification of subtracted cDNA (up to 30 μg) € 100 per 1 μg

Service pricing request on-line

 

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 2002-2011 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