Hi All,
I have just realised that at the end of the Ampure clean-up, I eluted with x100 Tris buffer by accident. Is this the source of my problems? And is there any way to solve this??
I have 120 human feacal samples which I am preparing for MiSeq at the moment. I am following the Illumina protocol and have the first PCR and clean-up completed (with very good PCR products).
I attempted to run the first plate with the Nextera library prep primers yesterday and when I ran the gel all I got back was a very faint band around where the first (16s amplicon) PCR product was (550bp) and no bands at all where they should be (630bp).
After rerunning 4 of my samples on a different PCR block and getting the same result, I gave a couple samples to my colleague who was doing a Nextera library prep and offered to try them. She got the same result and her prep didnt work.
Thank you!
I have just realised that at the end of the Ampure clean-up, I eluted with x100 Tris buffer by accident. Is this the source of my problems? And is there any way to solve this??
I have 120 human feacal samples which I am preparing for MiSeq at the moment. I am following the Illumina protocol and have the first PCR and clean-up completed (with very good PCR products).
I attempted to run the first plate with the Nextera library prep primers yesterday and when I ran the gel all I got back was a very faint band around where the first (16s amplicon) PCR product was (550bp) and no bands at all where they should be (630bp).
After rerunning 4 of my samples on a different PCR block and getting the same result, I gave a couple samples to my colleague who was doing a Nextera library prep and offered to try them. She got the same result and her prep didnt work.
Thank you!
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