Hi folks,
Sorry for putting very basic questions here. I am very new to NGS and I have planned to perform ITS1 amplicon sequencing on MiSeq. I have combed through several articles and found that 2x250 PE would be a suitable option for me. Anyway, I have some questions about indexing.
1. I have ca. 50 samples in which I would like to multiplex them. In this case, single or double index is better? And may you briefly explain differentations between the two systems or suggest a publication for further reading?
2. In case of double index, is it possible to use Nextera indices or other custom indices generated for bacterial 16s metagenomics for fungal ITS? I understand that they should be fine as long as I have both color channels in each base positions. Can you suggest if there is any I should be aware of?
Thank you in advance!
Sorry for putting very basic questions here. I am very new to NGS and I have planned to perform ITS1 amplicon sequencing on MiSeq. I have combed through several articles and found that 2x250 PE would be a suitable option for me. Anyway, I have some questions about indexing.
1. I have ca. 50 samples in which I would like to multiplex them. In this case, single or double index is better? And may you briefly explain differentations between the two systems or suggest a publication for further reading?
2. In case of double index, is it possible to use Nextera indices or other custom indices generated for bacterial 16s metagenomics for fungal ITS? I understand that they should be fine as long as I have both color channels in each base positions. Can you suggest if there is any I should be aware of?
Thank you in advance!
Comment