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DNA extraction - Salting Out: Difference between revisions
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DNA extraction - Salting Out (view source)
Revision as of 06:01, 26 November 2008
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==Notes== | ==Notes== | ||
*Other Salts (NaCl, AmOAc, LiCl) may also be used. See here for an explanation of how ethanol precipitation works and a description of the roles of different salts [http://bitesizebio.com/2007/12/04/the-basics-how-ethanol-precipitation-of-dna-and-rna-works/] | *Other Salts (NaCl, AmOAc, LiCl) may also be used. See here for an explanation of how ethanol precipitation works and a description of the roles of different salts [http://bitesizebio.com/2007/12/04/the-basics-how-ethanol-precipitation-of-dna-and-rna-works/] | ||
*I routinely rely on the denaturation step of Hot-start Taq in my PCR to take care killing proteinase K | *I routinely rely on the denaturation step of Hot-start Taq in my PCR to take care killing proteinase K (unless it's just not carrying over through the precipitations, either way it hasn't been a problem for me in PCR downstream). | ||
*I have successfully used this protocol to prepare quality PCR template from fish, mammal and insect tissue. | *I have successfully used this protocol to prepare quality PCR template from fish, mammal and insect tissue. | ||
*Works well for high throughput (e.g. 96 well plate extractions). To remove ethanol following precipitation/wash steps simply invert the plate over a sink then spin gently (~100 rcf) inverted over absorbant paper towelling. | *Works well for high throughput (e.g. 96 well plate extractions). To remove ethanol following precipitation/wash steps simply invert the plate over a sink then spin gently (~100 rcf) inverted over absorbant paper towelling. | ||
