IGEM:UNAM/2009/Notebook/IGEM project/2011/05/30

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Gel of PBBMRCS-5 plasmid
Abstract

Today I did a gel of the DNA plasmid PBBMRCS-5, the gel showed that the plasmid was cleaned of chromosome.


 * 1. I did the agar to 1%.
 * 2. I put the gel in running buffer and fill the wells of the gel.


 * 3. Volume of substances in each well of the gel:
 * 1st lane -> 5 μl of green ladder.
 * 2nd lane -> 5 μl of plasmidic DNA PBBRMCS5 + 2 μl of loading dye.
 * 3rd lane -> 5 μl of plasmidic DNA PBBRMCS5 + 2 μl of loading dye.
 * 4th lane -> 5 μl of plasmidic DNA PBBRMCS5 + 2 μl of loading dye.
 * 5th lane -> 5 μl of green ladder.


 * 4. I put the gel in the BIO-RAD during 110 minutes to 90 Volts.
 * 5. I washed the gel with distilled water.
 * 6. I put 100 μl of etide bromure miligramo/mililitro with distilled water in a tray and put the gel. Wait 10 minutes.
 * 7. I flushed the gel with water and put in distilled water  during 10 minutes.
 * 8. I saw the gel in the GBOX.

Note: PBBMRCS-5 was extracted using alcaline lysis and 15μL Plasmid DNA digested with XBA I and Hind III.

PBBRMCS-5 Plasmid Digestion

Leave all night at 37ºC in agitation.