UCSC In-Silico PCR
 
>chr5:177403762-177404289 528bp CCTAGGCTTCATGTCCCACTC AGAAGGTAGGGCACGGGTTC
CCTAGGCTTCATGTCCCACTCatgcccgcgcagccggcaccgccgaagcc
tcagcccacgacccggaccccgcctcagtcccagaccccgggaggttagg
aagtgggggggggaaggaggagccgagagggcgccgggcgagctagattc
cggccagccggccgcgggctctccgtcctcagcccctgctcctccacagc
cttgccggcgaagcgggagcagccgccttccctgaccaggaacggcccac
tgagctgcgggcagcggctccgcaagagtctgtcttcgatgacccgcgtc
gttggcgggctggtggcgctacgcggggcgcacccctacatcgccgcgct
gtactggggccacagtttctgcgccggcagcctcatcgccccctgctggg
tgctgacggccgctcactgcctgcaggaccggcgagtacccgcccgccca
gagccgccccaggggccgcggctcctccgtctcccagcgcagcttccacg
ctgcacccGAACCCGTGCCCTACCTTCT

Primer Melting Temperatures
  Forward: 61.0 C cctaggcttcatgtcccactc
Reverse: 62.6 C agaaggtagggcacgggttc
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