User:Paul Rothenberg/Notebook/Pauls Notebook/2013/02/25

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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