Isolation and preliminary characterization of cryptic satellite DNA in pea |
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Authors: | Lynne V.M. Wall John A. Bryant |
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Affiliation: | Department of Plant Science, University College, P.O. Box 78, Cardiff, CF1 1XL, U.K. |
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Abstract: | Optimum conditions have been established for isolation of ‘cryptic’ satellite DNA from the genome of pea (Pisum sativum), using gradients of CS2SO4 containing silver ions. At an Ag+ :DNA-P ratio (R) of 0.1, and at alkaline pH, four fractions are obtained: mainband (buoyant density 1.437 g cm3; 67% of total DNA), satellite I (buoyant density 1.582 g/cm3; 7% of total DNA), satellite II (buoyant density 1.520 g/cm3, 11% of total) and satellite III (buoyant density variable between 1.45 and 1.51 g/cm3; 15% of total). The reiterated DNA content of these four fractions has been investigated by reassociation experiments conducted over a Cot range of 1 × 10?5 to 2.0. All four fractions contain at least two kinetic components; each fraction, including the mainband, consists at least partly of highly reiterated DNA. Ribosomal RNA hybridizes only to the mainband. |
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Keywords: | Leguminosae pea repetitive DNA reassociation. |
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