UCSC In-Silico PCR
 
>chr5:177402068-177402557 490bp CGTGCCAGGTGAGCTCTTAG GTGGAGAAGCTGTGAATCGC
CGTGCCAGGTGAGCTCTTAGcccggttggcgcccttccccgaggccgtca
ggcacaaatctcaggtccacagcgctgagctgcgtgtttccgacccaggg
tgattccggaggcccgctggtgtgtgaggaccaagctgcagagcgccggc
tcaccctgcaaggcatcatcagctggggatcgggctgtggtgaccgcaac
aagccaggcgtctacaccgatgtggcctactacctggcctggatccggga
gcacaccgtttcctgattgctcagggactcatctttccctccttggtgat
tccgcagtgagagagtggctggggcatggaaggcaagattgtgtcccatt
cccccagtgcggccagctccgcgccaggatggcgcaggaactcaataaag
tgctttgaaaatgctgagaaggaaagctcttttcttcatgggtcccgccg
ggaaatgccaagacagaaaaGCGATTCACAGCTTCTCCAC

Primer Melting Temperatures
  Forward: 61.1 C cgtgccaggtgagctcttag
Reverse: 61.0 C gtggagaagctgtgaatcgc
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