Hello,

I am have been trying to use the IDT Alt R Genome Editing detection kit to assay some potential clones of KO cells I have generated. This kit is essentially a T7 endonuclease mismatch cleavage assay. I have been trying to use this kit for months and for the life of me I cannot get any products on my gel once I run it. I know that I have pure DNA product when I go in (I run a gel of my PCr products before continuing on with the mismatch assay), but when I image my gel at the very end, I have no product to speak of.

Also, the kit comes with two separate controls. One is a homo complex and the other is a heteromplex. These are subject to the T7 endonuclease treatment. The homocompelx is to supposed to result in a band at around 1000 bp and the heterocomplex should form two bands at 800 and 200 bp respectfully. I am able to see the homo complex on the gel, but for some reason I can never see the proper heterocomplex forming.

If anyone that has used this kit successfully can provided some tips for success that would be greatly appreciated. Thank yoU!