There's a minimum of assays to publish results from a mutation using TALEN?

[Alvaro Sequeida]

Hi everyone, I've mutated an exon of a gene from rainbow trout, but I want to know if there's any minimum of assays in order to publish my results. What I have done is a transcient transfection of a cell line (right and left arm of TALEN + GFP vector to check transfection efficiency), then I've got gDNA from the cells (pool of GFP+ cell and not transfected) and I've amplified the specific region that was mutated and I've digested the PCR product with a RE that has a recognition site in the spacer of the TALEN pair. Next I purified the digestion resistant band, and TA clone it, and then I pick different clones to sequence them and confirm the mutation. I've checked the mutation already, so the TALEN pair works just fine, but I want to know if it neccesary to put an mutation rate or something, I know that it can be done just comparing the bands from a gel, but I don't have the exact protocol. Oh, I also did a Western Blot, and the protein level is lower than compared to control cells. I don't know if it is necessary to do a qPCR to confirm that the gene is no longer expressed or something. I'll sort the cells using the GFP, but I want to do it to make another kind of assays with them. 

Thanks in advance for your advises !!