Does anyone have experience of co-feeding rrf-3 RNAi plasmid containing bacteria with a second RNAi containing bacteria ? If so, would they be willing to mail a pL440 plasmid with rrf-3 in it to me? (I am based in North America).
The reason is that I want to make a double mutant with a loss of function of one allele and a gain-of-function of a second allele, and wonder if it would work better in a RNAi supersensitive background. I could make the double gof;rrf-3 worm and use that in a feeding experiment, but just thought that including rrf-3 RNAi bacteria may be easier in the long run.
hi,
i made one, a derivative of my pYZT,which I generated a few years ago
in my graduate lab.
I should bring aliquot with me (check back soon)and you could contact
me by email; one friend has used it for a similar experiment as you.
@pYZT: itself a blue-white selection RNAi T vector,
amplifiable in host DH5alpha /HT115 and generate easily
RNAi T- vector DNAs with single enzyme digestion as you may know,
then t/a cloning your PCR products bearing A
overhang, white colonies are usually insert positive
[colony PCR can easily confirm it].published.
I made it for cloning “whole” target genes cDNAs library
generated by SSH, i.e.a target cDNA RNAi library. it works fine .
