Custom primers for NextSeq: will they work on MiSeq and HiSeq?

[rudzik79]

I am wondering whether my experiment (which was originally designed to work on a NextSeq) will work on MiSeq and HiSeq.

It is a library prepared with MIP technique and the PCR primers are

CAAGCAGAAGACGGCATACGAGATNNNNNNNNACACGCACGATCCGACGGTAGTGT
AATGATACGGCGACCACCGAGATCTACACATACGAGATCCGTAATCGGGAAGCTGAAG

My sequencing primers:

FOR CATACGAGATCCGTAATCGGGAAGCTGAAG C
REV ACACGCACGATCCGACGGTAGTGT A
INDX ACACTACCGTCGGATCGTGCGTGT A

I would be grateful for a quick response

Anna

[SNPsaurus]

We use custom primers and run libraries on NextSeq and HiSeq. MiSeq has a higher temperature for binding so a given primer may not work on MiSeq.

[rudzik79]

Thanks for your reply!
OK, now, how do I check that? What are the boundaries for Tm for MiSeq, NextSeq and HiSeq?