IUBio Biosequences .. Software .. Molbio soft .. Network News .. FTP

single embryo PCR

m.champagne at cellbio.duke.edu m.champagne at cellbio.duke.edu
Wed Jul 22 14:47:57 EST 1998


Hi Merav,

if you need to find out if your gene is taken out by a specific deficiency
stock, i would recommend doing the polytene squash technique in which
you hybridize a digoxygenin probe.  Even though our lab is quite good at
a variety of PCR techniques, it took me a while to figure how to get the
conditions right to get consistent product out of my PCR controls (low DNA
yield, false positives, false negatives...).

The end result you will get out of your single PCRs will not be as convincing
either, it will be a statistical decision (1/4 of all embryos you tested
shouldn't have your product...).  With the polytenes, the result is clear and
obvious, as long as your deficiency is big enough to be mapped physically
(ie:  has anyone defined the limits...  60E3-E7?).

Let me know what you think.

m

---------
In article <35B4BD4A.D73 at shum.huji.ac.il>, meravc at shum.huji.ac.il wrote:

> Does anybody happen to fortuitously have a reference to a protocol or 
> an article describing single embryo PCR used to map a gene to a 
> deficiency/deletion?
> 
> Thanks in advance for the help.
> 
> -Merav.
> 
> ---------------------------------------------------------------------
> 
> Merav Cohen
> Department of Genetics
> Hebrew University of Jerusalem
> Jerusalem, Israel
> 
> meravc at shum.huji.ac.il
> +972-02-6586038
> 
> for some cool drosophila biology, check out: 
> http://gamesdomain.uol.com.br/directd/mac/arcade/fly-dont-die.html



More information about the Dros mailing list

Send comments to us at biosci-help [At] net.bio.net