UCSC In-Silico PCR
 
>chr16:85913005+85913374 370bp TCTCCTGAGATAAAGGTGACAATATG CACCTCTCAGGGCTGAGCTAC
TCTCCTGAGATAAAGGTGACAATATGgttttacttacacatgaactgttc
atttttacaaacactggagccccaggtgggagatcagtgactgagctgcc
ctcctctccctcccctgactgtgcagccttctgtggacgattacatgggg
atgatcaaaaggagcccttccccgccggaggcctgtcggagtcagctcct
tccagactggtgggcgcagcagcccagcacaggtgagggtgggtggccta
gaattgtcaccaggcatggcctgaagggtttacggcattccaccagccag
ggacggagtgggggtggttgttgctatcatgatccatgtctcattaaagG
TAGCTCAGCCCTGAGAGGTG

Primer Melting Temperatures
  Forward: 59.9 C tctcctgagataaaggtgacaatatg
Reverse: 61.5 C cacctctcagggctgagctac
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.

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