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PHP scripts call R functions to check, insert and format data, and to perform online statistical analysis or visualisation.
10 μl PCRs with M13 forward and reverse primers were carried out to check insert size and the presence of a single insert and, from these, 96 colonies were chosen for 100 μl PCRs.
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Transformed bacteria were applied to LB plates containing 50 μg ml-1 ampicillin for selection and 40 μg ml-1 X-gal for detection of α-complementation [ 77]. White and positive colonies were picked for colony PCR screening to check inserts.
Most prominently, they disagree on the appropriateness of Southern Blotting as an exhaustive method to check the insert's stability and targeted comparative analyses as a method to detect potential hazards.
R [ 16] functions check and insert them into the database.
Then EcoRI was used to digest the isolated plasmids to check the insert size.
Plasmids were digested with EcoRI and XhoI to check the insert presence.
All sequences were searched for vector segments to check for insert integrity.
A subset of transformant colonies from each cloning reaction was analyzed by PCR in order to check the insert size.
Periodically during BAC library construction, pulsed-field gel electrophoresis analysis was used to check the insert size of randomly-selected NotI-digested BACs.
The strong hairpins formed by the mirrored pairs prevented sequencing through them, so their successful insertion was verified by checking the insert length via PCR and by the presence of an un-sequenceable hairpin.
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