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Old 04-22-2017, 01:56 PM   #5
Carcharodon
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Location: Honolulu, HI

Join Date: Jul 2015
Posts: 40
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I've heard of a lot of folks having good success with phenol-chloroform extractions. They report high yields of high-quality, high-weight gDNA.

One thing we like to do with the Qiagen Kits (DNeasy Blood and Tissue) is a triple-elution. We use 35uL water or buffer to elute some DNA, then 50uL water or buffer to elute more DNA, and finally do 2x 50uL elutions.

The first elution (35uL) tends to have most of the "sheared" DNA - those small fragments are washed out.

The second elution (50 uL) tends to have high-weight DNA, but may have some smaller/sheared fragments.

The third elution (100 uL) tends to have nothing but high-weight gDNA, though you may find the yield is lower than in the other two elutions.

If need be, you could combine the second and third elutions to achieve the desired mass of gDNA.

For the second two elutions (elution 2 and elution 3): you may consider warming your water/buffer to 70 degrees C.
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