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Old 11-04-2015, 10:20 PM   #1
Thuur
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Location: NL

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Default ultra long fragments capturing

Is it possible to capture ultra long fragments, for example >200kb. For this I need to extract High molecular weight DNA from Human blood. But then I want to capture these long fragments for example chromosome 6 short arm or a specific 200k fragment, Hopefully there is a technique without PCR.

Anybody has experience with this?
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Old 06-19-2019, 12:34 AM   #2
tangjj
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Hi,have you successfully captured ultra long fragments? Recently I want to capture a 200kb fragment from high molecular weight DNA. I wonder if you have any successful experience.
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Old 06-21-2019, 06:44 AM   #3
pmiguel
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Old school method involved embedding cells in agarose and lysing/prepping the DNA in situ. Initially these were just little slabs of agarose. But later moved more to higher surface area collects of agarose "beads".
Anyway, then they get run on a pulsed field gel for size selection.
Anyone using agarose for their large fragment preps these days?

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Phillip
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Old 06-27-2019, 10:05 AM   #4
JTH-Genome
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Smile BioNano

I agree with Phillip. Use the agarose plugs for the lysis. It might be old school, but it is making a comeback! You can use the BioNano protocol for high MW DNA. It's one of the best protocols around.

https://bionanogenomics.com/support-...isolation-kit/
https://bionanogenomics.com/products/bionano-prep-kits/

Or you can do your own homebrew version with agarose plugs.
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