I am new to NGS.
Goal is to find absolute abundance of yeast species in a mixed sample.
I plan to use PCR-free methods since PCR could cause amplification biased.
Plan is to use DNA fragmentase and size select ~300-400bp on Gel-> clean up->
end repair->A tail-> ligation/barcode/adaptor-> MiSeq 250 pair end.
From this millions of seq.(from different part of DNA) to figured out species and it's relative abundance??? Feels like i am missing some info. somewhere.
Any potential biased or problems?
above all this i do open for other complete new suggestions....
Goal is to find absolute abundance of yeast species in a mixed sample.
I plan to use PCR-free methods since PCR could cause amplification biased.
Plan is to use DNA fragmentase and size select ~300-400bp on Gel-> clean up->
end repair->A tail-> ligation/barcode/adaptor-> MiSeq 250 pair end.
From this millions of seq.(from different part of DNA) to figured out species and it's relative abundance??? Feels like i am missing some info. somewhere.
Any potential biased or problems?
above all this i do open for other complete new suggestions....