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DNA pellet


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18 replies to this topic

#16 HomeBrew

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Posted 15 March 2011 - 02:51 AM

I think that the problem is not that the "pellet" is not sticking to the tube, I think it's that the "pellet" is of low density and it floats. This makes me suspect that you actually have two "pellets" in your prep -- one DNA pellet which is pelleting at the bottom of the tube (and is likely not very visible), and another "pellet" of cell debris that is floating.

I'll bet you could fish this floating stuff out of the prep and proceed without it, and your PCRs would still work fine.

#17 Ameya P

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Posted 16 March 2011 - 10:03 PM

Probably, you have identified the problem HB.

I will try and look for the non-visible pellet, which makes the PCRs work and see, if I can discard the floating stuff, in a sample. The problem is that the phenomenon is quite inconsistent now. So, its difficult to predict where we will see floats and where we wont. Also, these aren't research samples which I could experiment with. At the end of the day, I have to give results with the samples I get.

but thanks anyways and I shall post new developments....

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#18 chimpsarehungry

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Posted 05 April 2011 - 07:53 AM

Do you want the pellet to stick just so you can get rid of the ethanol and then resuspend the pellet in TE? Because it does stick much better if you pipette the pellet and about a ml of ethanol out and put that into a 1.5ml tube and then centrifuge that for a few minutes then you can just pour off the ethanol and the rest will air dry quickly.

#19 Ameya P

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Posted 05 April 2011 - 09:43 PM

Do you want the pellet to stick just so you can get rid of the ethanol and then resuspend the pellet in TE? Because it does stick much better if you pipette the pellet and about a ml of ethanol out and put that into a 1.5ml tube and then centrifuge that for a few minutes then you can just pour off the ethanol and the rest will air dry quickly.


or I pipette out a little ethanol, so that i leave the pellet and ethanol in the same tube, spin it down and see if it sticks. Thanks for the advice, chimps :)

Actually, the person I was working with moved out of the lab and I have a new apprentice :P . No sticky issues with his extraction (so far)!

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