I have been trying to clone my amiRNA construct in 2 binary vectors since a whole year till now. I am using the amiRNA construct using Weigl group method, and have got my piece after long trials into the pENTR of gateway system, however have failed totally to clone it in the pEarleyGate100 or pEarleyGate101.
After digestion of pENTR I made PCR, and am sure my band is there but it seems that the problem occurs with ligation with the new plasmid. Any suggestions, or recommendation to better vectors, that will be expressed both in bacteria and plant.
Thanx and have a nice Day!
Edited by Nahola, 08 March 2012 - 01:52 AM.