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Team:University of Ottawa/15 July 2008
From 2008.igem.org
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| - | ==Today | + | ==Today in the Lab== |
'''Tammy''' | '''Tammy''' | ||
:'''Transformation of XL10 Competent E. coli cells with pDR197:AtCKX2''' | :'''Transformation of XL10 Competent E. coli cells with pDR197:AtCKX2''' | ||
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:'''Digestion of AtCRE''' | :'''Digestion of AtCRE''' | ||
::<li> AtCRE was digested again using the same protocols as above, except that it was run on the "digest and denature" program on the thermal cycler. It was left overnight. | ::<li> AtCRE was digested again using the same protocols as above, except that it was run on the "digest and denature" program on the thermal cycler. It was left overnight. | ||
| + | '''Dan''' | ||
| + | :'''Gel of overnight PCR''' | ||
| + | ::<li>Sample 0B did not show up on the gel, and was thus considered unsuccessful. This information was coherent with results from the absorbance measurements. | ||
| + | :'''PCR at 35 cycles''' | ||
| + | ::<li>PCR reaction of 0B was run overnight at 35 cycles instead of 29. | ||
Revision as of 14:44, 21 July 2008
Contents |
Today in the Lab
Tammy
- Transformation of XL10 Competent E. coli cells with pDR197:AtCKX2
- 1 uL Pure pDR197:AtCKX2
- 1 ul 1:10 dilution pDR197:AtCKX2
- 1 ul 1:100 dilution pDR197:AtCKX2
- 100 ul XL10 E.Coli per reaction
- 900 ul LB Broth per reaction
- 3 LB Ampicillin (50 ug/mL) Plates
- Began Incubation of transformed cells at 1:50 pm.
Matt
- Inoculation of 97 - BY4742 cells
- I inoculated the 97 BY4742 cells once again for glycerol stock tomorrow
- PCR confirmation
- Second PCR confirmation with F58, F59 was performed and gave the correct band but also gave another weird band that I don't know what it is.
- Digestion of PTP2
- A digestion was performed on the PTP2 amplification product to create sticky ends using BamHI and xhoI and I left it in the PCR machine overnight.
Chris
- Digestion of AtCRE
- AtCRE was digested using EagI at 37 C for about one hour
- the result was run on a 1% gel, which showed two bands, both of which were expected
- Gel Extraction of AtCRE
- the AtCRE was extracted from the gel using the gel extraction kit and protocol
- unfortunately, the final AtCRE sample was vacuumed up with the other waste by accident
- Digestion of AtCRE
- AtCRE was digested again using the same protocols as above, except that it was run on the "digest and denature" program on the thermal cycler. It was left overnight.
Dan
- Gel of overnight PCR
- Sample 0B did not show up on the gel, and was thus considered unsuccessful. This information was coherent with results from the absorbance measurements.
- PCR at 35 cycles
- PCR reaction of 0B was run overnight at 35 cycles instead of 29.
