Haynes Lab:Notebook/Engineering PC-TFs/2013/10/05: Difference between revisions
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* PEG/MgCl<sub>2</sub> | * PEG/MgCl<sub>2</sub> | ||
** Make 50 mL | ** Make 50 mL | ||
***Put water, then | ***Put water, then MgCl<sub>2</sub>, then PEG 8000 in 50 mL conical tube | ||
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<br> <br> <br> <br> <br> <br> <br> <br> | <br> <br> <br> <br> <br> <br> <br> <br> <br> <br> <br> | ||
*Perform restriction digest for CMV promoter | *Perform restriction digest for CMV promoter | ||
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| PstI|| 1.0 μL | | PstI|| 1.0 μL | ||
|- | |- | ||
| | | 10x FastDigest buffer || 3μL | ||
|- | |- | ||
| dH<sub>2</sub>O || | | dH<sub>2</sub>O || 15 μL | ||
|- | |- | ||
| || 30 μL --> 37°C/ ~10 min. | | || 30 μL --> 37°C/ ~10 min. |
Revision as of 21:06, 11 October 2013
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October 5, 2013
PEG/MgCl2 Precipitation
DNA Clean & Concentrator
Gel Electrophoresis
Higher molecular mass DNA precipitates at lower PEG concentrations than lower molecular mass DNA, according to Lis and Schleif 1975, NAR. I kept DNA from the supernatant and discarded the precipitated DNA. Therefore, I should see larger bands at 2% PEG and smaller bands at 15% PEG.
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