User:Paulina Alatriste/Notebook/UNAM Genomics Mexico 2011/2011/07/07

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Standard plasmid isolation

ABSTRACT

  • Isolation of pBBRMCS5 plasmid that we assume is already standardize.

  • The petri dish I made yesterday with the different colonies looks like this:

In this dish we only observe one red colony so we take some bacteria from that line and we put them in both liquid medium and solid medium. The red colony suggests that one ligation of the plasmid with the standard PCR was correct.

  • So the next step was to isolate this standard plasmid, for that Pablo and I extracted plasmid today using the following protocol but we did step 13 twice. After that we run an electrophoresis gel at 120V for 35 min.

Lanes:

1. Ladder 10 kb
2. Pablo´s isolation 1.
3. Pablo´s isolation 2.
4. Paulina's isolation 1.
5. Pauilna's isolation 2.

The isolation in the third lane doesn´t go well. The next step is to digest with EcoR1 and Pst1 in order to see if the region with the preffix and suffix was correctly added to the plasmid.



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