I got low clustering and very bad sequencing primer efficiency on NextSeq 500 run in my MIP project. The sequencing primers (read1, read2 and two index primers) are customized. We have tried either to spike our primer into illumina sequencing primer wells or add them into optional wells which are designed specifically for costumer specific primers. Both didn't give use any sequencing results. Do anyone have any suggestions ? thanks.
Does any know what is typical Tm for sequencing primers from illumina ? Looks like our customer specific primers don't anneal to reads very well. Is there anything that I can do to improve it in our Nextseq 500 run ? Thank you in advance.
Does any know what is typical Tm for sequencing primers from illumina ? Looks like our customer specific primers don't anneal to reads very well. Is there anything that I can do to improve it in our Nextseq 500 run ? Thank you in advance.
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