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Old 09-25-2017, 09:11 PM   #7
Meyana
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Location: Japan

Join Date: Sep 2017
Posts: 40
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Quote:
Originally Posted by torben View Post
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.
Thanks for this very useful info! I was also thinking of cutting everything in half, but was unsure as to whether this would be a bad choice somewhere in the procedure. Nice to hear other people are doing this successfully!
Now currently awaiting a new Qubit, so I can accurately measure my samples, not trusting the NanoDrop for these small concentrations...

I will ask around for the presence of a speed-vac, will probably need one no matter how I do this, as I am eluting my DNA in a too high volume from my Ampure beads (anyone know how little elution volume you can get away with using pr. vol Ampure?)
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