Haynes Lab:Notebook/Engineering PC-TFs/2013/09/26: Difference between revisions
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| PstI|| 1.0 μL | | PstI|| 1.0 μL | ||
|- | |- | ||
| Green 10x FastDigest buffer || | | Green 10x FastDigest buffer || 0μL | ||
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| dH<sub>2</sub>O || | | dH<sub>2</sub>O || 18 μL | ||
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| || 30 μL --> 37°C/ ~10 min. | | || 30 μL --> 37°C/ ~10 min. | ||
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''PEG/MgCl<sub>2</sub> Precipitation'' | ''PEG/MgCl<sub>2</sub> Precipitation'' | ||
* This procedure was adapated from [http://openwetware.org/wiki/Protocol_Size_selective_DNA_precipitation_by_PEG/MgCl2 Size selective DNA precipitation] | |||
*Measure the following into 1.5 mL tubes. | *Measure the following into 1.5 mL tubes. | ||
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* Wash the agarose gel flask. | * Wash the agarose gel flask. | ||
* Run gel for 45 min at 100 V. Check gel under UV light. | * Run gel for 45 min at 100 V. Check gel under UV light. | ||
[[Image:hiddendragonsofthetemple.jpg|250px|CMV PEG/MgCl<sub>2</sub> Precipitation 9/26/2013]] | |||
Higher molecular mass DNA precipitates at lower PEG concentrations than lower molecular mass DNA, according to [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC342844/ 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. | |||
Revision as of 12:22, 28 September 2013
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September 26, 2013Restriction Digest
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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