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DNA extraction - details about ethanol precipitation - (Sep/06/2005 )

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Chargeswitch from Invitrogen does not use high salt or alcohol

Precipitation is not required an9d the protocols are completely aqueous

-John Buckels-

u need a lot of DNA to see it by saltingout with EtOH. why not centrifuge it, even u dont see DNA thread u would get DNA, u may not see a ppt even, just dissolve in minimum of TE or H2O and run on 0.8% gell.


QUOTE (amty @ Sep 7 2005, 02:52 PM)
Iam extracting DNA from mosquitoes,
how much sample should I use?(currently using 10nos/ml)
I have followed ethanol precipitation
but I could not see the white thread
of DNA on adding ethanol.

To see the white thread, there has to be tons of DNA in the solution. If the amount of DNA is too little to see the white thread, you add a small amount of glycogen (maybe 2ul or so) and centrifuge it. By adding the glycogen, you will see a white precipitate, which is the glycogen and the DNA. The glycogen will dissolve in TE or even water/Tris.Cl at pH8 and they don't interfere with most DNA manipulations, except for sequencing which requires extremely clean DNA.


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