TF transposon genotyping PCRs for granulosa differentiation

For the transposon genotyping PCRs, I used NEB Q5 polymerase, following the manufacturer's recommended protocol for PCRs from genomic DNA. I used 20 sec. extension time at 72 °C, 35 cycles, with the following primers and conditions:

Gene Fwd Rev Anneal °C Expected bp
NR5A1 CATCCTGGTGAAAGACGCTC AGCGTAATCTGGAACATCGTATGGGTA 67 277
GATA4 GCCCTCAGACCTCCAGCAAG AGCGTAATCTGGAACATCGTATGGGTA 70 131
FOXL2 CCTGCAGTTCGCTTGTGCC AGCGTAATCTGGAACATCGTATGGGTA 70 159
TCF21 ACCCGGTCAACCTGACGTG AGCGTAATCTGGAACATCGTATGGGTA 70 158
RUNX1 CGGGACCGATCCAGCCTACC CGTGGACGTCTCTAGAAGGA 67 148
RUNX2 CGGGACCGATCCAGCCTACC TGCTTGCAGCCTTAAATGACT 66 198

Then I ran the PCR products on a 2% agarose gel with TAE buffer and ethidium bromide.