Quote:
Originally Posted by ZAAB
Seems to me that one of the biggest 'deterrents' of contamination between samples within one prep, between preps across time, etc [U]is to use enough template DNA so that any contamination won't be preferentially amplified
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I agree - it is a numbers game. The issue however is that some applications are setup to find rare variants (e.g. circulating cancer cells or low concentrations of microbes in a complex mixture) - in these scenarios a low level (and unpredictable) baseline of contamination can be problematic. Again, (to sound like a broken record) the workflow - be it 1-step and 2-step - depends on end-use of the data.
The point I am trying to make here is that contamination in NGS workflows is real and needs to be managed - never more so than when amplifying amplified DNA.