Gel of PBBMRCS-5 plasmid
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
|NEBuffer 2 10X
|PBBRMCS-5 Plasmid DNA
Leave all night at 37ºC in agitation.