Hello everyone,
i am conducting microbiota sequecing using MiSeq.
When i was doing index pcr according the manual (http://support.illumina.com/content/dam/illumina-support/documents/documentation/chemistry_documentation/16s/16s-metagenomic-library-prep-guide-15044223-b.pdf).
i wrongly set the PCR program one cycle more as follows:
95 for 3min
-95 for 30s
I don't think it will matter. Call Illumina, they have a really good tech support, that will give you some piece of mind.
Do you mean you carried out 9 cycles? If so I'm sure it will be fine and you should be checking the insert size and DNA quantity before taking the libraries any further anyway.
You may have a higher library concentration than expected but could dilute to the correct concentration.
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