Scaling up Reclone distribution - DNA preps

Hi All

We have a bottleneck on DNA preps of the Open DNA Collections and I know that is also affecting the Reclone Hubs and Nodes - not everyone has the facilities to prep hundreds of constructs.

A colleague pointed me to this company for DNA prep services and they are very competitive: https://www.biozilla.bio/plasmid-dna-uk

We still need to do an initial miniprep of everything and supply >1 ug DNA so we can’t send immediately because there are some recent parts that my lab have which are still only glycerol stocks. However, they advertise midiprep/Petiteprep yields (0.2 mg to 0.5 mg) for £17 per sample (minipreps of 5-10 ug are £3).

iGEM only ship 1-2ng DNA per well, my lab were adding much more to our initial distributions because our automated magbead preps were low quality and had a lot of RNA contamination.

If we get a high quality prep of 0.2 mg DNA, we can make 100,000 distribution plates and probably not need to prep again for a long time, at least for distribution. Obviously for downstream cloning the end user would need to amplify and prep.

I’m tempted to do this and then up to 100 Reclone Hubs/Nodes can receive enough DNA to make an additional 1000 plates before they need to prep any of their own DNA. Midipreps of up to 500 parts would be £8500, then of course there are plates and shipping.

I’m going to include this budget an upcoming grant proposal, if anyone else gets the chance to do similar then please do, so we maximise chances!

Also, other ideas or tips on service providers are very welcome.

Jenny

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Hi Jenny.

Here we use the MiraPrep protocol, that gives a much higher yield of high quality DNA using the MiniPrep kit. The protocol is very much the same, we only include a wash with 98% ethanol. Please see our protocol here: MiraPrep protocol
The reference: The Miraprep: A Protocol that Uses a Miniprep Kit and Provides Maxiprep Yields

It might be useful, at least one prep would be enough for many plates.

Natalia

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