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Old 09-25-2017, 01:41 AM   #6
torben
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Location: Norway

Join Date: Oct 2012
Posts: 20
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Quote:
Originally Posted by Meyana View Post
Perfect, thank you so much!
Do you have any experience in starting with under the recommended 1ng DNA for the Nextera XT kit? My starting material is a small brain tissue sample, so I am worried about how much material will be available for me. I will test next week when all my reagents are delivered, but I really doubt I will reach 1ng mtDNA...
The easiest way to scale down is to reduce all the volumes in the reaction. I usually use half the volumes given in Illumina's protocol, and I have collaborators who only use a quarter of the volume. You need to have your DNA in a pretty small volume though (≤1.25-2.5 Ál). I elute my DNA in pure water and use a speed-vac to reduce the volume, but be carefull not to overdry the samples. Alternatively, I guess you can also use the normal volumes and only reduce the Tagment Mix to matvh your DNA input but I've never tried this.
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