Nick Gervais
posted 8 months ago
Ph.D. student developing CRISPR-based technologies and studying fungal pathogenesis and drug resistance in the Shapiro Lab at the University of Guelph.

Resuspending Twist Oligo Pools?

Genetic EngineeringMolecular BiologyGeneticsMicrobiology Twist Oligo Pools Twist Bioscience Oligo Pool

Hi everyone,

I've received a custom oligo pool from Twist Bioscience and they provided 70ng of it. We talked to one of the reps and they said not to resuspend it to any less than 10ng/μL, which means I'll be adding less than 7μL of buffer, which is obviously not a lot. I was wondering if anyone had any practical tips for how to make this work and not lose any DNA. Is 10ng/μL really the lowest you can safely store DNA at? What centrifugation time/speed do you use to spin it down first? Do you heat it at all before reuspending? Do you vortex or just pipette up and down to mix? Any advice would help! Thank you.

Guy Rohkin8 months ago

Seems like an odd proposition considering that DNA is highly stable. Do you know how much of these oligos you need for downstream reactions? 7uL does seem like a small volume and you can run the risk of it evaporating (and thus increasing the remaining concentration) or just overusing it.

Guy Rohkin8 months ago

My experience has been with the lyophilized oligos from IDT and they are usually suspended in pretty big volumes based on the molarity. Wouldn't make sense to me why at a certain concentration it is "safer" per sé. I used to just add nuclease-free water and pipette up and down to mix it to get all those little particles in.

Nick Gervais8 months ago

@Guy Rohkin okay glad I'm not the only one who thinks that's a little strange. I only need 10ng for my cloning reaction but I would feel way more comfortable resuspending it in a larger volume and then just using a larger volume of it during cloning...

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