Hello Everybody,
I have extracted genomic DNA from 7 different bacterial species and pooled into a single solution (1ng from each ). After library preparation from the pooled DNA and sequenced by Illumina HiSeq-2000, I can,t find three of the seven bacterial species in sequencing results. what can be the possible reason?
Thanks
MJ
Hello, it's not very clear what you did, sorry if I am wrong, but maybe you should make a PCR 1 separately for each sample, clean it and only then pool them together for metabarcoding? As I understood you pooled it after extraction.
Do you know that the quality of the DNA from all 7 species was comparable? Sometimes you might have a degraded DNA prep that gives you plenty of OD reading but is not a good substrate to build the library. So it may be something that simple.
any one can tell me how much is the maximum number of DNA sample that could be pool together for sequencing
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