Detection of simple sequence length polymorphisms by silver staining

Detection of simple sequence length polymorphisms by silver staining M . KLINKICHT a n d D. T A U T Z Zoologisches Institut der Uniuersitrft Munchen, Luisenstr. 14, D-8OOO Miinchen 2, Germany Analysis of simple sequence length polymorphisms (SSLP; zlso called microsatellites) is a powerful tool for paternity determinations and population screening (Tautz 1990). The technique normally involves PCRamplification with 32P-labelledprimers, resolution of the resulting bands on acrylamide gels and detection by autoradiography (Tautz 1989). However, because it is usually desirable to avoid radioactive compounds, we have tested silver staining of the polyacrylamide gels as an alternative. The use of silver staining has been described before for the analysis of PCR products (Allen, Graves & Budowle 1989). We show here in addition that the required two nucleotide resolution for the analysis of SSLPs can easily be obtained on standard gel sizes. Gels were poured in a vertical gel apparatus (14 x 16 cm; 0.75 mm thick). The gels are made up to be the same as sequencing gels, but without urea. The polyaaylamide concentration may vary between 6 1 0 % and is usually 8%.A sample (2-5 pl) of a standard PCR (Saiki et al. 1988) reaction is loaded and the samples are run either for 3 http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Molecular Ecology Wiley

Detection of simple sequence length polymorphisms by silver staining

Molecular Ecology, Volume 1 (2) – Aug 1, 1992

Loading next page...
 
/lp/wiley/detection-of-simple-sequence-length-polymorphisms-by-silver-staining-wVkzkvg0Vk
Publisher
Wiley
Copyright
"Copyright © 1992 Wiley Subscription Services, Inc., A Wiley Company"
ISSN
0962-1083
eISSN
1365-294X
D.O.I.
10.1111/j.1365-294X.1992.tb00166.x
Publisher site
See Article on Publisher Site

Abstract

M . KLINKICHT a n d D. T A U T Z Zoologisches Institut der Uniuersitrft Munchen, Luisenstr. 14, D-8OOO Miinchen 2, Germany Analysis of simple sequence length polymorphisms (SSLP; zlso called microsatellites) is a powerful tool for paternity determinations and population screening (Tautz 1990). The technique normally involves PCRamplification with 32P-labelledprimers, resolution of the resulting bands on acrylamide gels and detection by autoradiography (Tautz 1989). However, because it is usually desirable to avoid radioactive compounds, we have tested silver staining of the polyacrylamide gels as an alternative. The use of silver staining has been described before for the analysis of PCR products (Allen, Graves & Budowle 1989). We show here in addition that the required two nucleotide resolution for the analysis of SSLPs can easily be obtained on standard gel sizes. Gels were poured in a vertical gel apparatus (14 x 16 cm; 0.75 mm thick). The gels are made up to be the same as sequencing gels, but without urea. The polyaaylamide concentration may vary between 6 1 0 % and is usually 8%.A sample (2-5 pl) of a standard PCR (Saiki et al. 1988) reaction is loaded and the samples are run either for 3

Journal

Molecular EcologyWiley

Published: Aug 1, 1992

Keywords: ; ;

References

You’re reading a free preview. Subscribe to read the entire article.


DeepDyve is your
personal research library

It’s your single place to instantly
discover and read the research
that matters to you.

Enjoy affordable access to
over 18 million articles from more than
15,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

Search

Query the DeepDyve database, plus search all of PubMed and Google Scholar seamlessly

Organize

Save any article or search result from DeepDyve, PubMed, and Google Scholar... all in one place.

Access

Get unlimited, online access to over 18 million full-text articles from more than 15,000 scientific journals.

Your journals are on DeepDyve

Read from thousands of the leading scholarly journals from SpringerNature, Elsevier, Wiley-Blackwell, Oxford University Press and more.

All the latest content is available, no embargo periods.

See the journals in your area

DeepDyve

Freelancer

DeepDyve

Pro

Price

FREE

$49/month
$360/year

Save searches from
Google Scholar,
PubMed

Create folders to
organize your research

Export folders, citations

Read DeepDyve articles

Abstract access only

Unlimited access to over
18 million full-text articles

Print

20 pages / month

PDF Discount

20% off