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R. Saiki, D. Gelfand, S. Stoffel, S. Scharf, R. Higuchi, G. Horn, K. Mullis, H. Erlich (1988)
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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
Molecular Ecology – Wiley
Published: Aug 1, 1992
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