[Chief_Lazy_Bison]

Hello!

I recently tried to use some homemade Nextera Index PCR primers (0.5uM final concentration in a limited cycle index PCR reaction). The bioanalyzer results for my first PCR generation looked great. However, after the 2nd PCR I saw bands which were larger, smaller, and the correct size for the desired product. Any advice on how to optimize reactions for homemade Nextera index PCR primers?

[Kaizen]

Correct, we have tried IDT synthesized oligos and they worked! No need for Nextera XT index kit from illumina anymore.

Hi kcchan,
Could you please share your oligo sequences? I've tried paired end multiplexing of 384 samples with custom oligos from Sigma and some of them (100 oligos) seem to not work as good as the Illumina's.