UCSC In-Silico PCR
 
>chr5:35874060+35874836 777bp GATGTCTTTGGGACTGGAAC GCCATGAATGATGGCTTCTG
GATGTCTTTGGGACTGGAACtgagaatgtgtaggaactgagacatctcct
ccctaaaatttgcaacaggggtgaacatccctctcatcatctcctgctct
ggcttcttttccttggtagaaagtcaagaagggaagagagcattggtacc
tttgatgctagatcacgtttacatttcaagtggcagatgctctgggcctg
gtcacccaagtcaatgccttttaaaccaaaatccctccataaagctgtca
aatatgtctcttaactgaaaagcaactttcaggaaataataagtgggccc
acattactaagtaaatgcaaagcaccctgagaccctacccccactgcatg
gctactgaatgctcaccacaatctattcttgctttccaggggagatggat
cctatcttactaaccatcagcattttgagttttttctctgtcgctctgtt
ggtcatcttggcctgtgtgttatggaaaaaaaggtgaccttcttcaacta
ataaagagggtgattgtgtgggatcacggacagtcagagcttaagcccca
tttattgatgagaaaaccacaaaggggattaaggcatttcacgaatttag
tgcccagtatccctatctatcctcagcgaatttccacagttaatttcata
agaggcaaaaagatattaactggacattaggcaaacgctgtccccaaagt
aagaattccgtaatgcaatgtttcccaagcttttcttccagtatctccta
aaggttgCAGAAGCCATCATTCATGGC

Primer Melting Temperatures
  Forward: 57.0 C gatgtctttgggactggaac
Reverse: 61.6 C gccatgaatgatggcttctg
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