IGEM:UNAM LCG/2009/Notebook/Hydrobium etli/2011/09/08
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I ran a gel to calculate the concentration of the 20 Anderson collection promoters and the pBBR1MCS-5 backcbone in order to have the information to do the ligation (to insert the promoters in the backbone). The promoters were ok but the Backcbone did not appear in the gel, I will need more backbone (more concentrated).
I ran 2 gels because I needed more lines than the number of lines that you can put in a single gel.
1. Ladder 2. J23100 3. J23101 4. J23102 5. J23103 6. J23104 7. J23105 8. J23106
1. Ladder 2. J23107 3. J23108 4. J23109 5. J23110 6. J23111 7. J23112 8. J23113 9. J23114 10. J23115 11. J23116 12. J23117 13. J23118 14. J23119 15. empty 16. backbone pBBR1MCS-5
In this gene everything appear to go well, I just have to calculate the concentrations. The part that appear less concentrated is J23104 which I already expected. gel 03-sept
This second gel had more problems. First I forgot to charge the plasmid backbone, so I did it when the gel has already ran 10 minutes (so the backbone only ran 30 min). I knew that its size was aprox 4.8 Kb so I was only interested in knowing the concentration, but at the end nothing appeard in its line, I suspect that it is necessary to have it more concentrated, Helena will extract it again. And then I do not have the number of lines I was expecting, the plasmids should be from line 2 to 14 and the line 14 is empty, probably I loaded the part J23107 and the ladder in the first line, or maybe I never loaded J23119. Anyway I will have to run again a gel to see the concentration of the new plasmid backbone extraction.