UCSC In-Silico PCR
 
>chrX:9440337+9440743 407bp CCTCATCCACGCCAGAATCGGTAAA AATGATCGCCTGCAATCTTCCCACA
CCTCATCCACGCCAGAATCGGTAAAatgcgtacatgaagatgcagcagac
aatgcatccaatcacaaataggatccaatcctcacgaacacgccgatcga
tgacttgaagtgtcgagttagagagacccaactcaaaatatgtttttaaa
agttgaaattagaaatgtaacttacagcttggccaatttcatgcattttt
ctcccaacgccgcgtaaatttaaatgctgggatttcaagttttcgagtac
agcagatccttgacttatcagatcgtcaagctgcaattattgcttttaaa
ttcatttttttcaattaacggaagacaaaaatttaattttaagtggtaag
taccataaacgttatctgaaaactcacatgagTGTGGGAAGATTGCAGGC
GATCATT

Primer Melting Temperatures
  Forward: 70.7 C cctcatccacgccagaatcggtaaa
Reverse: 71.4 C aatgatcgcctgcaatcttcccaca
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