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is it possible to extract DNA from filter paper by using autoclaved water? - DNA extraction from filter paper by water (May/10/2006 )

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Hi

i got a plasmid from a fellow on filter paper. i have cut the filter paper into small bits and have put autoclaved water to it. the filter size is of thumb size. i have added 10 ul of water and have left it in fridge for 2 days. i checked today with spec but it seems the dna is not yet extracted from the paper. that is real s@*%:-(

please help me with sopme way to get the DNA out of the paper which is already soaked in water..

thanx all for help and support

debashih

-debashish-

Forget the spec -- you've probably got plenty. Try transforming some competent cells.

-HomeBrew-

the spec comes out to be negative. that makes me skeptical to go for cell transfection. i am not pretty sure if i have DNA at all in the tube...

-debashish-

hi
if you let water and paper during 2days, DNA has eluted. i don't know how many water you added, but the conc is not enough concentrated to get a good spectrum.
But that doesn't mean you don't have DNA. Transform bacterias and do a prep.

-fred_33-

i added 100microliter of water for a small bits of blot paper cut from a thumb sized one. i will try to get it transformed in bacts

-debashish-

use 10┬Ál in order to be sure.

-fred_33-

I recently got some plasmids on a filter paper. I used 100uL of water and left it in for 2 days. I transformed 1ul of the elute and got plenty of colonies.

Good luck

-scolix-

do i need to leave the filter paper in water for two days at room temp or in fridge?? i left in the fridge.

-debashish-

when I rec'd a plasmid that way, I left it in ~50ul 10mM tris for about an hour, and then transformed 2ul and got loads of colonies

-aimikins-

QUOTE (debashish @ May 10 2006, 08:10 PM)
the spec comes out to be negative. that makes me skeptical to go for cell transfection. i am not pretty sure if i have DNA at all in the tube...

Forget the OD reading.
You can get a plenty of colonies on your plate with a very low OD reading.

For me, 50-100 ul water will be added, leave it for 1-2 days and then take ~10 ul for transformation.

-carassius-

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