UCSC In-Silico PCR
 
>chr5:177403994-177404563 570bp ACCTACCGGAACGTGACTGC CGGGTCATCGAAGACAGACTC
ACCTACCGGAACGTGACTGCcgagcaagcgcggaactggggactgggcgg
ccacgccttctgccggtgcgccgcgtggggctgggtgacccctccgcccc
agggctccgggctcccggcgctctaacggcgccccgtcgtgtggctacag
gaacccggacaacgacatccgcccgtggtgcttcgtgctgaaccgcgacc
ggctgagctgggagtactgcgacctggcacagtgccagaccccaacccag
gcggcgcctccgaccccggtgtcccctaggcttcatgtcccactcatgcc
cgcgcagccggcaccgccgaagcctcagcccacgacccggaccccgcctc
agtcccagaccccgggaggttaggaagtgggggggggaaggaggagccga
gagggcgccgggcgagctagattccggccagccggccgcgggctctccgt
cctcagcccctgctcctccacagccttgccggcgaagcgggagcagccgc
cttccctgaccaggaacggcccactgagctgcgggcagcggctccgcaaG
AGTCTGTCTTCGATGACCCG

Primer Melting Temperatures
  Forward: 62.4 C acctaccggaacgtgactgc
Reverse: 62.2 C cgggtcatcgaagacagactc
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