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No colony after ligation? - (May/24/2006 )

Hello everybody, im doing ligation using insert:vector 3:1 and 1:1 but get nothing.
I dephosphorylated my vector pcDNA3.1(+).
What do you people think "dephosphorylation can cause problem?


I have not heard dephosphorilation of vector couses any problem,
maybe there is problem with your competent cells
check them.


I dont think dephosphorylation can cause any problem with liagtion unless both the vector and insert are dephosphorylated.


thanks to all for quick reply, I got colonies after changing competent cells as well as increasing my I:V ratio upto 1:3.
I got total of 14 colonies and should i go for prep of all?