is it possible to extract DNA from filter paper by using autoclaved water? - DNA extraction from filter paper by water (May/10/2006 )
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
Forget the spec -- you've probably got plenty. Try transforming some competent cells.
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...
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.
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
use 10µl in order to be sure.
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
do i need to leave the filter paper in water for two days at room temp or in fridge?? i left in the fridge.
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
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.