A rapid DNA preparation for PCR fromChlamydomonas reinhardtii andArabidopsis thaliana |
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| Authors: | Deborah A. Berthold Barbara A. Best Richard Malkin |
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| Affiliation: | (1) Department of Plant Biology and Integrative Biology, University of California, 94720 Berkeley, CA, USA;(2) Present address: Department of Biology, Colby College, 04901 Waterville, ME, USA |
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| Abstract: | A method for preparing DNA for PCR has been adapted from the forensic work of Walsh et al. (Biotechniques 10:506–513) for
use withChlamydomonas reinhardtii andArabidopsis thaliana. The method consists of a short incubation of cells or tissue in ethanol, followed by addition of Chelex-100 and heat treatment.
Following centrifugation, the supernatant is added directly to the PCR reaction; forChlamydomonas, amplification product is visible over a range of four orders of magnitude of starting cells. Using this method, DNA suitable
for PCR template can also be obtained fromArabidopsis leaf tissue without grinding, organic extraction or precipitation steps. This method may prove to be useful for other plant
and algal species. |
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| Keywords: | Arabidopsis thaliana
DNA isolation
Chlamydomonas reinhardtii
PCR amplification |
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