Term
| how many ml of bacterial (what kind?) culture were put in the centrifuge tube? |
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Definition
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Term
| what room was the refrigerated centrifuge in? |
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Definition
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Term
| how many rpms does the refrigerated centrifuge spin at? |
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Definition
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Term
| how many minutes is it centrifuged? (both times) |
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Definition
10 the first time
5 the second |
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Term
| what are the layers the first time it is centrifuged? |
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Definition
| E. coli pellet at bottom, nutrient broth at top |
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Term
| what are the layers the second time it is centrifuged? |
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Definition
| aqueous DNA on top, white protein "junk" in between, phenol and lipids on bottom |
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Term
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Definition
-2% sucrose solution to be isotonic to E. coli and keep the cells from blowing apart in centrifuge
-buffer to control pH
-ETDA - compound that stops enzymes from damaging DNA |
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Term
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Definition
-detergent breaks apart cell membrane (binds to lipids)
-NaCl - Na+ balances the negative charge of DNA |
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Term
| what is the detergent used? |
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Definition
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Term
| how long is the centrifuge tube kept in the water bath? what temp? why? |
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Definition
10 minutes
60 degrees Celsius
speeds up process of breaking apart cell membrane |
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Term
| how many ml of phenol is added? |
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Definition
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Term
| which compound is not soluble by phenol? |
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Definition
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Term
| how many volumes of ethanol is added to the aqueous DNA solution? |
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Definition
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Term
| what is used to move the aqueous DNA solution out of the 1st centrifuge tube to the 2nd? |
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Definition
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