L4440 digestion

Model Organism: Worms Methods & Reagents
1

Hi there

So i tried T7 primer PCR to confirm my gene of interest in L4440 RNAi. But no PCR product was obtained.Now i am trying to do a restriction digest to on the ahringer plasmids. But how to decide on which enzymes for digestions.

2

I assume you’ve looked at the sequence and restriction map available though Addgene?

3

So i can not use enzymes between 52-165, i guess (as this region is used for TA cloning by EcoRV digestion). I am just new to this plasmid and trying to figure out, how they cloned the gene of interest (RNAi clone) and how to confirm these plasmids with digestions as the T7-PCR did not work.