I want to clone ZAP about 2100bp. I am facing difficulties to clone this gene can anyone recommend me a best method and vector to clone. I want to use this clone for A549 cell line
I used PCMV5a and pNL vector but failed to get ligation i used fusion kit (Clone jet )also used T vector to get successful clone but fail all the times
Use PCR to amplify the coding sequence with both 5' and 3' sequences overlapping to your vector of interest (about 30-40 bases) and then use Gibson assembly mix. It always works for me!