UCSC In-Silico PCR
 
>chr5:35856798+35857161 364bp CTGTTTCTGGAACTTGCCTTC ATTCAAACCCAGTGCCTGAC
CTGTTTCTGGAACTTGCCTTCttttttgctgtgtttatacttcccttgtc
tgtggttagataagtataaagccctagatctaagcttctctgtcttcctc
cctccctcccttcctcttactctcattcatttcatacacactggctcaca
catctactctctctctctatctctctcagaatgacaattctaggtacaac
ttttggcatggttttttctttacttcaagtcgtttctggagaaagtggct
atgctcaaaatggtgagtcatttctaagttttcttatggattttggatta
tctgtagcatggtttcaggttattcagttccctaacagacctgaGTCAGG
CACTGGGTTTGAAT

Primer Melting Temperatures
  Forward: 59.0 C ctgtttctggaacttgccttc
Reverse: 60.0 C attcaaacccagtgcctgac
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