UCSC In-Silico PCR
 
>chr16:85921029+85921602 574bp TGGCTAGGGTCAAAGACAGTG GTCTTTGGAAATGATAGTTAACCG
TGGCTAGGGTCAAAGACAGTGcccaccccctttggagcctccgcctctgc
ctctgactttctgcacctcccatctagattgagcagctgtatgtccggca
actggcagaagaggctgggaagagctgtggagccggctctgtgatgcagg
cccccgaggagccgccgccagaccaggtcttccggatgtttccagatatt
tgtgcctcacaccagagatcatttttcagagaaaaccaacagatcaccgt
ctaagtgcgtcgcttgggcgccccaccccgtctgcgtcctgcatccatct
ccctgttacagtggcccgcatcatgattaaagaatgtggatccctctgtc
tggggtgggatgccttactttgcacttaatttaataagggcattctcgga
ggagtagacgtttaatacgaagtggcggcatagccctgccgagatgtcgg
tgatggcctggatgctgtaaccacaacctgtggctaaaaattttattttc
tatcctttacccgtcattatcattagttgctatgattctttctgcatttt
CGGTTAACTATCATTTCCAAAGAC

Primer Melting Temperatures
  Forward: 60.3 C tggctagggtcaaagacagtg
Reverse: 58.2 C gtctttggaaatgatagttaaccg
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