syringe method for DNA recovery

Paul N Hengen pnh at fcs260c2.ncifcrf.gov
Mon Jun 6 17:23:07 EST 1994


      ** Bouced e-mail directed to osp086 at clvax.bangor.ac.uk **
                      Re:  PCR product from gel

> Paul,
> Regarding your recent posting on this subject on methods & reagents:
> I noted with interest the remarkable yield using the syringe method and so
> gleefully sent off for a reprint of the biotechniques paper which I 
> received quite promptly this morning. However the authors refer to some
> kind of "allergy syringe" which they suggest may be quite important due to
> its low dead space volume, and I have no idea what type of instrument this
> is. I do not know of a UK supplier for the particular NJ firm that provided
> said syringes so in order to buy correctly I could do with a bit more info.
> Is this just a disposable plastic 1ml syringe or am I being too hopeful. If
> in the unlikely event that it is what guage size is the needle? Finally
> I don't know which companies supply the US and the UK but if you know of any
> that do and which supply these syringes could you let me know.
> Sorry to burden this upon you but it saves on air mail and I don't know Dr Li's
> e-mail.
> 
> Many thanks,
> Craig Wilding
> School Of Ocean Sciences,
> Menai Bridge,
> Gwynedd,
> Wales,
> U.K.
> LL59 5EY.


Dear Craig:

I had to post here because your e-mail address is causing my message to bounce.
Li and Ownby report in their paper that they used a Becton-Dickinson 1 ml 5540
allergy syringe.  Becton-Dickinson & Co. is located in Rutherford, NJ 07070
USA.  For the experiment that I described in my posting where I got the best
yield of DNA fragments from 250 ul of agarose, I used a Becton-Dickinson 3 cc
syringe (Reorder no. 9585) stuffed with siliconized polyallomer fluff. This
syringe does not come with a needle attached and I used it without one.  To get
the highest recovery, I used a pipetman to withdraw the last little bit of
liquid from the end of the syringe where there is some dead space.

I've also used a Becton-Dickinson 1/2 cc U-100 Insulin syringe (Reorder no.
9461) that comes with a 28 1/2 Micro-Fine IV needle for smaller volumes of
agarose (up to ~100 ul) in the same way. This syringe comes with the needle
attached, but I removed it with a pair of pliers so that I could get the
maximum recovery. There is not as much dead space at the tip of this one and I
think that for small volumes like 50-100 ul of gel slice, you could use any
tuberculin or insulin type disposable plastic syringe without the needle.

Good Luck and Happy Squeezing :-)

-Paul.

*******************************************************************************
* Paul N. Hengen, Ph.D.                           /--------------------------/*
* National Cancer Institute                       |Internet: pnh at ncifcrf.gov |*
* Laboratory of Mathematical Biology              |   Phone: (301) 846-5581  |*
* Frederick Cancer Research and Development Center|     FAX: (301) 846-5598  |*
* Frederick, Maryland 21702-1201 USA              /--------------------------/*
*******************************************************************************



More information about the Methods mailing list