hi everyone. For GBS-analyses i need high quantity and quality gDNA of around 400 samples of a violet species (dried material). When i´m using the Qiagen DNeasy plant kit 96 and elute with 50ul TE i get between 10 and 30 ng/ul measured with a nanodrop. However as the nanodrop often seems to overestimate DNA concentration, my yields might be to small for the analyses (optimum 50 - 100ng/ul).
To optimice the DNA concentration i thought of two different ways ...
1) making two parallel DNA extractions of the same sample, elute each with 100 or 200ul TE, combine them and finally make an ethanol precipitation.
2) making two parallel DNA extractions of the same sample, elute the first with 50 or 100ul TE, use the resulting elution from the first sample again for the second sample and in this way increase the final concentration.
In the moment i prefer variant 2) as it is easier, much faster and includes less steps than variant 1) (-> 400 ethanol precipitations in single 2ml tubes). What do you think, is this worth doing or do you have any other suggestions how to concentrate the DNA in the 96 wells?
Thanks for any answers,
Ben
To optimice the DNA concentration i thought of two different ways ...
1) making two parallel DNA extractions of the same sample, elute each with 100 or 200ul TE, combine them and finally make an ethanol precipitation.
2) making two parallel DNA extractions of the same sample, elute the first with 50 or 100ul TE, use the resulting elution from the first sample again for the second sample and in this way increase the final concentration.
In the moment i prefer variant 2) as it is easier, much faster and includes less steps than variant 1) (-> 400 ethanol precipitations in single 2ml tubes). What do you think, is this worth doing or do you have any other suggestions how to concentrate the DNA in the 96 wells?
Thanks for any answers,
Ben
Comment