Abstract
The products of sequencing reactions are separated on thin, low percentage (usually 6%) polyacrylamide gels. Normally, these gels are 40–50 cm in length, 20 cm wide, and between 0.3 and 0.4 mm thick. Longer gels (to enable more sequence to be read from a single run) up to 100 cm in length can be poured, as can wider gels (to enable more clones to be loaded onto a single gel). However, these larger gels are more difficult to handle after electrophoresis, that IS, during fixing, drying, and autoradiography. The methods given in this chapter deal only with the “standard“ size sequencing gels, but the principles are the same for larger gels, except that more gel mix will be needed, and different sizes of combs, spacers, and glass plates may be needed
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Biggin, M. D., Gibson, T J., and Hong, G. F. (1983) Buffer gradient gels and 35S label as an aid to raprd DNA sequence determination Proc. Natl. Acad. Sci USA 80, 3963–3965
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© 1993 Humana Press Inc. Totowa, New Jersey
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Littlebury, P. (1993). Pouring Linear and Buffer-Gradient Sequencing Gels. In: Griffin, H.G., Griffin, A.M. (eds) DNA Sequencing Protocols. Methods in Molecular Biology™, vol 23. Humana Press. https://doi.org/10.1385/0-89603-248-5:115
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DOI: https://doi.org/10.1385/0-89603-248-5:115
Publisher Name: Humana Press
Print ISBN: 978-0-89603-248-4
Online ISBN: 978-1-59259-510-5
eBook Packages: Springer Protocols