UCSC In-Silico PCR
 
>chr5:35867248+35867540 293bp CCACTGCATACAGGAACTCCTAC AACCTACAGAATGTCCAGACACAG
CCACTGCATACAGGAACTCCTACctgaatcaagacatatcccctttttat
tcctacagtggggccctcgtggaggtaaagtgcctgaatttcaggaaact
acaagagatatatttcatcgagacaaagaaattcttactgattggaaaga
gcaatatatgtgtgaaggttggagaaaagagtctaacctgcaaaaaaata
gacctaaccactataggtaagaagttgtatataaaagtatggttgtcact
tttgggctacctgaaaacaCTGTGTCTGGACATTCTGTAGGTT

Primer Melting Temperatures
  Forward: 60.1 C ccactgcatacaggaactcctac
Reverse: 59.6 C aacctacagaatgtccagacacag
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