How to remove small amount of plasmid from filter paper - (Sep/19/2005 )
I received a plasmid that I requested from an overseas lab and they shipped it to me on a piece of filter paper. There is only 140 ng of plasmid spotted on the paper. Does anyone have any suggestions on how to remove such a small amount of plasmid and recover enough that I can carry on with a transformation? Thanks........
go to whatman's website and look for "clonesaver"
they will tell you how to get plasmids off filter paper
apparently this is the new thing in plasmid storage. i have only used it a couple of times but so far, so good...doesn't seem like it should be so easy but it is
Cut out the area of the filter paper containing the DNA and stick in a 1.5 ml eppendorf with about 500 ul of TE buffer. Swirl it around with a pipet tip and then use about 1.5-2 ul for transformation of supercompetent e. coli. This works everytime.
PS, Save the tube in case in transformation fails the first time.
this is how I do it,
cut out the circled area, put it in 100 or 200microlitre of TE. keep it at 37 degree for 4hrs.Take 10 microlitre of it and transform highly competent cells.
save the paper and the elutant.
Save the four hours, do it right away, it works every time.
The DGRC website has instructions on how to transform clones that they send you on Whatman paper. There is something in the paper that is toxic to the cells, so they suggest washing in TE first and then putting the cells directly on the paper disk in the tube. I've tried it a couple of ways and even have used the wash to transform cells. The long and short of it is that you only need one good clone from the filter, right? Just resuspend in a little TE, add cells and heatshock. Should work fine.
The DGRC protocol is here:
FTA Disc Protocol
I recently got DNA on filter paper and was instructed to just cut out a small piece containing DNA and put it directly into competent cells and transform as usual. It worked great! I have also just used water to extract the DNA and transformed from that, and have been equally as successful.