IGEM:UNAM LCG/2009/Notebook/Hydrobium etli/2011/09/19

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pBBR1MCS-5 plasmid extraction

Abstract

Vladimir helped me by making a plasmid pBBR1MCS-5 extraction from 4 different colonies: colony 1, colony 2, colony DH5α, colony S17. I ran a gel to see if the extraction were OK. I conclude that the extraction was succesfully done, the plasmid from S17 was the most concentrated.

Details

I ran the gel with this characteristics:

  • 120 V
  • 35 min
  • 4μL template + 3.5μL dye each lane.

Gel image

Lanes:

 1. ladder 500bp
 2. not interesting for this study
 3. not interesting for this study
 4. pBBR1MCS-5 1
 5. pBBR1MCS-5 2
 6. pBBR1MCS-5 DH5α
 7. pBBR1MCS-5 S17
 8. empty

Image:PBBR1MCS_extraccion.JPG

Gel analysis

All the marks are visible and in the correct place (between 4000 and 5000 bp) so I conclude that the extractions were good. The plasmid that is more concentrated is the one extracted from S17 (7th lane). The marks that are visible a little bit up the main marks are isoforms (concatenations for example).

Notes

The tubes are in my rack, the next step is to make a PCR from this parts to extract the plasmid backbone.



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