UCSC In-Silico PCR
 
>chr5:177404400-177404746 347bp AGGAGGGAGAGTGCGGAAAG CGTTGTCCGGGTTCCTGTAG
AGGAGGGAGAGTGCGGAAAGcagatgagagggaggcaggagagcccagcc
ttggctgcccagggagccccctttctcctcagacaccaaggcaagctgct
atgatggccgcgggctcagctaccgcggcctggccaggaccacgctctcg
ggtgcgccctgtcagccgtgggcctcggaggccacctaccggaacgtgac
tgccgagcaagcgcggaactggggactgggcggccacgccttctgccggt
gcgccgcgtggggctgggtgacccctccgccccagggctccgggctcccg
gcgctctaacggcgccccgtcgtgtggCTACAGGAACCCGGACAACG

Primer Melting Temperatures
  Forward: 63.1 C aggagggagagtgcggaaag
Reverse: 62.8 C cgttgtccgggttcctgtag
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