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Team:University of Lethbridge/Notebook/Project1July
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===July 3, 2008=== | ===July 3, 2008=== | ||
====Nathan Puhl, Munima, Christa==== | ====Nathan Puhl, Munima, Christa==== | ||
| - | No colonies on either plate but the DH5-alpha cells transformed with pSB1A7 were successfully transformed (~1500 cfu). There are two possibilities: 1. | + | No colonies on either plate, but the max-efficiency DH5-alpha cells transformed with pSB1A7 were successfully transformed (~1500 cfu). There are two possibilities: 1. the cells are competent but are low efficiency and we did not add enough DNA, or 2. The cells are not competent. To assess possibility one we will try more DNA. If that does not work we will try a protocol using electroporation. |
Revision as of 05:19, 4 July 2008
Contents |
July 1, 2008
Nathan Puhl
Subcultured RP1616 into liquid LB and pTopp cells into liquid LB + amp
July 2, 2008
Nathan Puhl, Christa, Munima
Plasma mini prepped pTopp cells
Attempted to make RP1616 cells competent using the open wet ware [http://openwetware.org/wiki/Preparing_chemically_competent_cells protocol]. 100 uL aliquots of cells were froxen in liquid nitrogen and stored in the -80 freezer.
Ran CheZ in pUC19 plasmid and pUC19 plasmid from last year with high range ladder. The pUC19 plasmid seems weird, I don't think it should have two bands; I will have to ask someone about that. The CheZ plasmid appears to be the right size (2600 + 620 = 3220 bp)
July 3, 2008
Nathan Puhl, Munima, Christa
No colonies on either plate, but the max-efficiency DH5-alpha cells transformed with pSB1A7 were successfully transformed (~1500 cfu). There are two possibilities: 1. the cells are competent but are low efficiency and we did not add enough DNA, or 2. The cells are not competent. To assess possibility one we will try more DNA. If that does not work we will try a protocol using electroporation.
