Sequencing a RNAi-Construct! - A question! (Jul/28/2005 )
in our laboratory everyone is telling me that there is no a chance to sequence my RNAi construct wich is already cloned in pART27. They say that the construct will form a hairpinstructure while being sequenced! Is that true? I received the construct from a former scientist and have to check it because I can't find reduced RNA-levels of the specific gene in my tranformated plant!
Another problem is that noone has the sequence of the pART27 vector! Does anyone has it, or knowes where to get it?
Thx for help!
That is NOT true.
I have sequenced many of my hairpin siRNAs cloned into a plasmid vector without any problem. Actually you have to sequence it before using it for transfection just to confirm the sequence is right.
hairpin structure does not occurs between denaturation step and elongation step during sequencing... The information you get is false...
I've succesfully sequenced siRNA without any problem.