Biology Forum › Molecular Biology › cDNA integration into Plasmid
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AuthorPosts
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October 18, 2006 at 5:25 pm #6039
Ultrashogun
ParticipantHow is a fragment of cDNA integrated into a plasmid although it does not have complementary extremities due to originating from restrictive endonucleasic digestion?
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October 18, 2006 at 7:18 pm #56934
oppox
ParticipantIve done a lab where we created a cDNA library I think but it was a long time ago and I dont remember the details so here is a link to something similar.
http://courses.washington.edu/dmandoli/ … ibrary.pdf
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October 18, 2006 at 9:36 pm #56936
canalon
ParticipantIf you used a PCR step with Taq polymerase, TA cloning is an option. Blunt end cloning is always possible, although the results are not very good.
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October 18, 2006 at 9:56 pm #56937
Ultrashogun
Participantquote oppox:Ive done a lab where we created a cDNA library I think but it was a long time ago and I dont remember the details so here is a link to something similar.http://courses.washington.edu/dmandoli/ … ibrary.pdf
Thank you, that gave me the answer, which is:
add linkers to all the cDNAs. These linkers are short oligonucleotide pieces of
dsDNA that contain a restriction site within them. Add the enzyme DNA ligase,
which will ligate the linkers to the ends of the cDNAs.
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