Dear B Ram, try following following method. make a very small hole at the bottom of a 0.5 ml PCR tube and cover it with some sterile glass wool. Put your cut gel band in this tube and close it. Now put this tube into a 1.5 ml eppendorf tube. using an appropriate balancing tube spin at 12,000 rpm for 5 min. you should get 70-80% of your DNA in the eppendorf tube which can be precipitated and resuspended in small volume. Alternatively, put this tube into 1.5 ml tube containing ~100 microlitre TAE buffer. Also add same amount of this buffer in 0.5 ml tube. Take two small pieces of platinum wires, dip one through space between 0.5 ml and 1.5 ml tube so that it immerses in the buffer. other piece of platinum wire should be purt in 0.5 ml tube buffer. apply low current (~ 10V) with black electrode connected to wire in 0.5 ml tube and red electrode to other one immersed in 1.5 ml tube buffer. This will make a small electrophoresis coloumn. after around 20 min, whole of DNA will move into buffer of 1.5 ml tube. Disacrd 0.5 ml tube along with its contents. precipitate DNA from buffer of 1.5 ml tube using sod acetate and cold ethanol. All the best.
On 12/25/10, [email protected] <[email protected]> wrote: > Send Methods mailing list submissions to > [email protected] > > To subscribe or unsubscribe via the World Wide Web, visit > http://www.bio.net/biomail/listinfo/methods > or, via email, send a message with subject or body 'help' to > [email protected] > > You can reach the person managing the list at > [email protected] > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Methods digest..." > > > Today's Topics: > > 1. gel elution of 6KB PCR product (B.Rama chandran) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Sat, 25 Dec 2010 18:52:35 +0530 > From: "B.Rama chandran" <[email protected]> > Subject: gel elution of 6KB PCR product > To: [email protected] > Message-ID: > <[email protected]> > Content-Type: text/plain; charset=UTF-8 > > Dear All, > > I am facing problem in the gel elution of 6KB PCR product, I am > getting very less yield (~10ng/micto liter). I am using Sigma gel > extraction kit (catalog no:NA1111). Since yield is very less I couldn't use > that product for restriction digestion. If you have any suggestion please > give me. If you know any other technique which will give better yield please > let me know that. > > regards, > B.Ram. > > > ------------------------------ > > _______________________________________________ > Methods mailing list > [email protected] > http://www.bio.net/biomail/listinfo/methods > > End of Methods Digest, Vol 67, Issue 8 > ************************************** > -- Dr V K Gupta Sr Microbiologist (Mol Biology) IMBL, Department of Entomology Pun. Agric. Univ., Ludhiana (Pb)-141004- India M: 081465-55515 _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
