UCSC In-Silico PCR
 
>chr5:177405258-177405871 614bp GACTGTGATAGGGCAGGGTG ATTCTGTAGGCCCAGGGTTG
GACTGTGATAGGGCAGGGTGggccaggcccctgggtagagcagggaagcc
ttgtctctttctacaggtgtgctaccacccccaactttgatcaggaccag
cgatggggatactgtttggagcccaagaaagtgaaaggtgctacacacag
cctctggggtggcctggggctctctcctcccgcctcattactctcctggt
atcaccagaccccacacacctgggattctggacccagccccttctctccc
tccacaataccctttggaagtccagagggagagttctgggaaggagtggt
cccattttgcaggtgggtaaaccaagcttggaaacttggagtagcaaggt
cacaaggcaagtaggttcaagaagggccttggcccccagctgtgtgactc
agctccctgctcttccttccaccatgtccatctctcagaccactgcagca
aacacagcccctgccagaaaggagggacctgtgtgaacatgccaagcggc
ccccactgtctctgtccacaacacctcactggaaaccactgccagaaagg
tgaggagatgtggaggacctgggcggggtgctgggggacaggggCAACCC
TGGGCCTACAGAAT

Primer Melting Temperatures
  Forward: 60.5 C gactgtgatagggcagggtg
Reverse: 61.3 C attctgtaggcccagggttg
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