User:Paul Rothenberg/Notebook/Pauls Notebook/2013/02/25: Difference between revisions
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== | ==25 February 2013== | ||
* | * Objectives: | ||
*1. Create a 1% agarose gel (1g agarose in 100 mL TAE) for gel electrophoresis | |||
*2. Run a DNA sample to test for expression | |||
*Procedure: | |||
*1. 50x TAE stock buffer was diluted to 1x TAE buffer, using 2 mL of 50x buffer and 98 mL of ultra-pure water (1) | |||
*2. 0.25 grams of agarose was dissolved in 25 mL of TAE | |||
*3. The TAE was microwaved for 45 seconds to dissolve the agarose | |||
*4. The TAE and agarose was poured into the electrophoresis chamber and left to harden; electrophoresis buffer was added following | |||
*5. 5 microliters of DNA and 1 microliter of dye; 6 microliters of the DNA ladder were added to the wells | |||
*6. The gel was run for 55 minutes at 90V | |||
*7. The gel was then placed in Ethidium Bromide rinse and shaken for 15 minutes | |||
*8. The gel was removed and placed in a TAE rinse and shaken for another 15 minutes | |||
*9. The gel was exposed in an electrophoresis chamber | |||
*Data: | |||
*1. No experimental DNA bands were visible under UV radiation. The gel will either have to be stained in ethidium bromide for a longer period of time, or a new gel will have to be rerun. | |||
*References: | |||
*1.http://openwetware.org/wiki/1X_TAE | |||
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25 February 2013
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