UCSC In-Silico PCR
  
>chrX:154563893+154564110 218bp ACACTAGTCCAGCCAAGGAGC CAACAGGAAGGTCGAACTGTG
ACACTAGTCCAGCCAAGGAGCtttacagaaatgcgtggcttgactggacg
gtttctgtttccaaaggatcgaggacatgaggaagcggcatgtcgaggtc
tcccaggcccccttgccccccgcccctggtgagtgagcgagaactgggcc
tgcgggaggaggtgggtggggagggcaggtgctgcgccgcgggaggtCAC
AGTTCGACCTTCCTGTTG
>chrX:154640327-154640544 218bp ACACTAGTCCAGCCAAGGAGC CAACAGGAAGGTCGAACTGTG
ACACTAGTCCAGCCAAGGAGCtttacagaaatgcgtggcttgactggacg
gtttctgtttccaaaggatcgaggacatgaggaagcggcatgtcgaggtc
tcccaggcccccttgccccccgcccctggtgagtgagcgagaactgggcc
tgcgggaggaggtgggtggggagggcaggtgctgcgccgcgggaggtCAC
AGTTCGACCTTCCTGTTG

Primer Melting Temperatures
  Forward: 60.8 C acactagtccagccaaggagc
Reverse: 60.7 C caacaggaaggtcgaactgtg
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, the formula by Rychlik W, Spencer WJ and Rhoads RE NAR 1990, which can be activated in Primer3 with PRIMER_TM_FORMULA=0.

Help
  What is chr_alt & chr_fix?
Replicating in-Silico PCR results on local machine