I am trying to ligate Il-8 and pIRES Ds Red,but I didnt succed.
I PCR IL-8 (which comes in a plasmid from Addegene).I designed primers with two different restriction sites,XhoI and Sac II.
I extracted IL-8 from the gel and digested with these 2 enzimes.I digested pIRES as well,with the 2 enzimes.
After ligation with DNA T4 ligase,and tranformation I didn`t get any colony.
I would like to know how I can precipitate my ligation before transformation.
And another question,should I use CAP?
Thanks in advance!!!
Edited by Bego, 12 April 2010 - 10:09 PM.