UCSC In-Silico PCR
 
>chr16:50716476+50716848 373bp AGCACAGATGCTGGCACTTC TTGGACAGTGAGCACCAAATC
AGCACAGATGCTGGCACTTCagggatgaatgaaagtctttttgggggatt
tgtagatttttttcttgtcttactagctccattttcaaatgtatttattt
tgtctctttagtttgcgcgataacaatatctcagaccgaggcatctgcaa
gctcattgaatgtgctcttcactgcgagcaattgcagaagttagcgtaag
tcagcctgggctgtggacaatgggctccaagtgccctggtctcaccccag
gtcgtgcagcctgggaagctgtgagtgatgggctggggcaggggctgttt
gcatgatggggggtgcaggtgattcctgcccagaggggaagggcaaccct
ggGATTTGGTGCTCACTGTCCAA

Primer Melting Temperatures
  Forward: 61.6 C agcacagatgctggcacttc
Reverse: 61.1 C ttggacagtgagcaccaaatc
The temperature calculations are done assuming 50 mM salt and 50 nM annealing oligo concentration. The code to calculate the melting temp comes from Primer3, the formula by Rychlik W, Spencer WJ and Rhoads RE NAR 1990, which can be activated in Primer3 with PRIMER_TM_FORMULA=0.

Help
  What is chr_alt & chr_fix?
Replicating in-Silico PCR results on local machine