08 December 2010 Dear All,
Below is a summary of answers I obtained for my questions on “Blotting paper for plasmid DNA”. Thank you all very much for your quick response and thanks to CCP4BB. -Partha ______________ Hideaki Moriyama: How about FTA filter papers (Whatman). https://dgrc.cgb.indiana.edu/vectors/ http://www.whatman.com/FTAElute.aspx ____________ Dima Klenchin : 1. what type of blotting/filter paper should be used? will any type of Whatman paper is good for this purpose? where (which supplier / catalog number) could I purchase that blogging paper? Just about any will do. Whatman #1 works fine, Whatman 3MM is nice because it is thicker and takes up more solution. 2. what is the typical lowest volume for spotting (will 5 micro lit. sufficient?) Any piece that can be conveniently fit in the eppendorf eventually - enough to make it wet. 5 ul is plenty for about 0.3 cm2 of Whatman #1. 3. Are these "spotted DNA" are good option for longtime storage (for about 6 months)? Yes as long as there was EDTA in the DNA solution (e.g., TE). Kept in the dark, of course. ___________ Patrick J. Loll: Whatman #1 is fine. We frequently use ca. 2 uL of a maxi-prep solution (so, say, maybe 100-200 ng). I haven't conducted any analysis of long term stability, but I wouldn't suggest storing it any longer than necessary (say, a week or so). If you need to store longer, there are better ways to do it. __________________ Konstantin v. Korotkov: I planning to spot plasmid DNA on a blotting paper for transport. I do not have any idea what type of blotting paper is used for this purpose. 1. what type of blotting/filter paper should be used? will any type of Whatman paper is good for this purpose? where (which supplier / catalog number) could I purchase that blogging paper? Any filter paper will do. I fact, any "paper" can be used as well. If you are worried about contamination, you could use sterile strips. 2. what is the typical lowest volume for spotting (will 5 micro lit. sufficient?) Even less will be sufficient. 1-2ul of a miniprep of plasmid DNA is plenty. 3. Are these "spotted DNA" are good option for longtime storage (for about 6 months)? I do not have long-term experience myself, but I would guess it should be quite stable as long as it's dry. _________________ Dhirendra K Simanshu: I can suggest at least what works for me. 1. I feel any clean Whatman filter/blotting paper would be fine. I have used 1mm for my purpose. Main purpose is to absorb all the liquid. 2. It will depend on DNA concentration. But I think 5-6 ul should be more than sufficient. 3. I am sure they will be good for six months in dried form. Generally I cut small pieces of Whatman Filter paper and then write the name of the construct using pencil in the corner and then drop 5-6 ul in the center and then draw a circle around the drop. Because once the drop is dried, this will help in locating the position. Generally I cover the paper with thin plastic wrap for transfer purposes. For extracting, I just cut the circled part of paper and then put it in few ul of Tris buffer at 37 C and then I have enough to transform and make large quantity of it.
