PCR Cloning
Sample sheet
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RESULTS
PCR products up to 3 kb will be cloned into a standard pCR-TOPO vector using TA
ligation. For efficient cloning, PCR products need to be 3’ adenylated and purified. Please
indicate on the sample sheet whether we need to perform these additional steps for you.
Positive clones containing inserts of the indicated length will be identified, and inserts of
3 clones will be verified by sequencing using vector-sepcifiek primers (productnr. BC660). Send in a minimum of 100ng of PCR product containing one specific band and
include a gel picture. Minimal fragment length for PCR products is 100bp for efficient
purification. Please note that the purified products should be free of Taq-activity,
nucleotides and primer-dimers, and that the product is 3’ adenylated (Proof reading
enzymes possessing 3’-5’ exonuclease activity produce blunt-ended PCR products, and
should be purified and tailed prior to cloning). BaseClear will only use organisms and
synthesis DNA that are classified under our GMO license. For this reason we require basic
sequence information to perform a risk analysis: donor organism, pathogenicity class,
gene name, and gene function (if known).
See also http://www.vrom.nl/ggo-vergunningverlening
Baseclear is entitled to check the sequence for contents, confidentiality guaranteed and
can reject any sequence that is not classified under our GMO license.
Please review the requirements for shipping of samples on our website
www.baseclear.com.
TERMS OF DELIVERY
Three clones containing inserts of the indicated length will be sequenced with vector
primers. Sequence delivery time for TA cloning is approximately 2 weeks. At additional
costs, a small aliquot of the isolated clones can be sent back to the customer as purified
DNA. BaseClear guarantees the complete confidentiality of all client information, project
data and biological samples during or after the termination of the contract. Please see
our website (www.baseclear.com) for more information.
I accept the general terms and conditions of
BaseClear*
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: ______________________
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: ______________________
Date
:__ / __/ ____
To be filled out by BaseClear:
Project Code
: __________________
Date of registration : _ _ / _ _ / _ _ _ _
*Our General Terms and Conditions apply to all projects BaseClear carries out and can be downloaded from our
website (www.baseclear.com/labservices/terms_conditions).
Version: 2013-01
F089-SYNTH-06
Page 1 of 2
PCR Cloning
Sample sheet
No.
Sample
name
Donor
organism
Potentially
hazardous?
(Y/N)
1
Size of PCR
product
(bp)
Conc.
(ng/ul)
Purification
needed?
(Y/N)1
A-tailing
required
(Y/N)1
Fragment
isolation
requested?
(Y/N)1
Additional
clones1
Return
purified DNA
(Y/N)1
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
PCR purification, productnr. BC-202; A-tailing, productnr. BC-663; Fragment isolation from gel, productnr. BC-206; Additional clones, productnr.
BC-661, Return of purified clone DNA to customer, productnr. BC-662.
1
Version: 2013-01
F089-SYNTH-06
Page 2 of 2