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Team:University of Chicago/Notebook/Shorelandhall
From 2008.igem.org
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'''Robert McConeghy's Notebook | '''Robert McConeghy's Notebook | ||
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| + | Thursday, August 28 | ||
| + | Prepared 1 L SOB medium using protocol from the White Notebook. This will be used tomorrow to make electrocompetent BL21 cells. | ||
Friday Aug 15 | Friday Aug 15 | ||
| - | The control worked (21 colonies), but the IGEM transformation failed. Harper and I discussed possible reasons for this failure. Nanodrop does NOT work for the IGEM DNA solutions because the TE buffer is clear whilst the DNA solutions are dyed from the spot. The nanodrop then gives a negative reading for concentration! | + | The control worked (21 colonies), but the IGEM transformation failed. Harper and I discussed possible reasons for this failure. Nanodrop does NOT work for the IGEM DNA solutions because the TE buffer is clear whilst the DNA solutions are dyed from the spot. The nanodrop then gives a negative reading for concentration! With 21 colonies, the competency of TOP10 cells turns out to be 10^5 colonies/mg. Oh snap. |
Thursday Aug 14 | Thursday Aug 14 | ||
Latest revision as of 20:15, 28 August 2008
Robert McConeghy's Notebook
Thursday, August 28 Prepared 1 L SOB medium using protocol from the White Notebook. This will be used tomorrow to make electrocompetent BL21 cells.
Friday Aug 15 The control worked (21 colonies), but the IGEM transformation failed. Harper and I discussed possible reasons for this failure. Nanodrop does NOT work for the IGEM DNA solutions because the TE buffer is clear whilst the DNA solutions are dyed from the spot. The nanodrop then gives a negative reading for concentration! With 21 colonies, the competency of TOP10 cells turns out to be 10^5 colonies/mg. Oh snap.
Thursday Aug 14 Transformed DH5a and TOP10 from freezer with IGEM DNA spot 7B which is a florescent protein. For a control, transformed TOP10 with pGreen.
Wednesday Aug 13 Chris's transformation did not work. How can this be?!?! Only the IGEM DNA must be the cause for this. We are unsure as to why the IGEM DNA can't help transformation.
Tuesday, August 12 Transformation didn't work. Turns out I used the wrong buffer solution. D'oh. Chris showed me how to use the electroporater. Using his cells and harper's plates, we tried to transform the IGEM DNA into cells using electroporation.
Monday, August 11 Today, I transformed my DH5a cells with some pGreen. For controls, I used some of Chris's cells...I think they were like HT155 or something. I tried pGreen in those and also the IGEM plasmid.
