I am in great problem and i need the help of you people.
I am trying to clone but it failed.
I am using insert of 1.4kb of psedomonas aergonisa and using pPICZalphaA as a vector which is 3.6kb, I got the correct pcr band, and when I double digested my vector and insert i had the correst band on the restriction dgestion, performed ligation , and tranformed into E.coli competent cell. got colonies , when i did the midi prep, and alkaline lysis method, and when determined the concentration of DNA i had good DNA concentration but when I tried to run on agarose no bands were visible.
As my tenure of thesis has finished and I am asked to stop at this point..
I dont know the reason why it did not work, it would be really nice of some people who could help me in this...
I am attaching the complete information of my experiment
Kindly reply and suggest me what reason can i give for the failure of my project
thanks and regards