UCSC In-Silico PCR
 
>chr5:35873199+35873819 621bp AAGTGTGGAAAGGGAGATCC CACAAAAGGCGCAAAGACAG
AAGTGTGGAAAGGGAGATCCtgggcagtgcctaactcacctgaataagac
ccatcatttcactctcctccttgaccactcacaacatcctttataagctc
agattctgtccctaattttgctgttgactcctttacgtatcagagctcct
tattctaacaaatacgagacaacttcagagaatgcttatgggactaaagg
aatcccaattgaaatgatttgggagatttaggcaacacctcttttcccat
cctaagaatgtaactgcactctactctctagcatgtgaatttatccagca
caaagctgacactcctgcagagaaagctccaaccggcagcaatgtatgag
attaaagttcgatccatccctgatcactattttaaaggcttctggagtga
atggagtccaagttattacttcagaactccagagatcaataatagctcag
gtaaggaatggtggtagagtttttgttccctcagagtgctttgcatgtca
aagtgtgggagcaagtgagaggaagattgttgaaactaacctgcaaaata
ggacacccttggagggcactcttacactttctttggagaatgacttgcct
gCTGTCTTTGCGCCTTTTGTG

Primer Melting Temperatures
  Forward: 58.0 C aagtgtggaaagggagatcc
Reverse: 61.9 C cacaaaaggcgcaaagacag
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