Biology Forum › Molecular Biology › cDNA integration into Plasmid
- AuthorPosts
- October 18, 2006 at 5:25 pm #6039
UltrashogunParticipantHow is a fragment of cDNA integrated into a plasmid although it does not have complementary extremities due to originating from restrictive endonucleasic digestion?
- October 18, 2006 at 7:18 pm #56934oppoxParticipant
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
- October 18, 2006 at 9:36 pm #56936canalonParticipant
If 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.
- October 18, 2006 at 9:56 pm #56937UltrashogunParticipantquote 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.
- AuthorPosts
You must be logged in to reply to this topic.
