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Old 02-09-2015, 09:56 AM   #1
Vinn
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Location: Sweden

Join Date: Nov 2014
Posts: 18
Default DNA extraction for amplicon sequencing

Hi folks,

Sorry for having a very basic question here. I am going to use amplicon sequencing to study endophytes in tree. I was wondering whether it is necessary to perform DNA extraction under laminar flow cabinet?

I know that it is important to prevent contaminations from airborne spores, especially the primer pair I use is universal one (ITS1F-ITS4). However, it is not practical to perform the extraction under LAF -- not mentioning about fungal spores that would contaminate in liquid nitrogen and etc. Do you have any opinions/suggestions?

Also would it help to reduce contamination and distortion of the results if I run PCR at low number of cycles (e.g. 20) instead of 35?

Thank you very much in advance!
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