So, i am an intern i need help understanding why we are using certain primers for my PCR. We are the SPACR gene at 645 bp. We did RNA isolation from transfected cells and after doing the "RT" part of RT PCR we are doing the PCR part. The people i am working with had me set up four different PCR reaction with 4 different sets of primers. The first set:323-342 R with 91-112 F.
The second: 879-900 R with 651-670 F.
Third: 1-26 Sal F with 620-45XBa R
Fourth : GAPDH F with GAPDH R.
i know that we used them to see what was really transfected but i don't understand what each one of them does or will tell me by using it.