Team:University of Ottawa/14 July 2008
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==Today in the Lab== | ==Today in the Lab== | ||
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::<li> the result was run on a 1% gel, resulting in five bands where only two were expected. | ::<li> the result was run on a 1% gel, resulting in five bands where only two were expected. | ||
::<li> using Vector NTI, we figured out that the incorrect enzyme was used for the digestion; in fact, EagI should have been used instead. | ::<li> using Vector NTI, we figured out that the incorrect enzyme was used for the digestion; in fact, EagI should have been used instead. | ||
+ | '''Dan''' | ||
+ | :'''PCR amplification of construct 1''' | ||
+ | ::<li> The amplification product was run on a gel however no clear bands showed up on the kodac machine. However the correct band was seen on the cutting UV machine. | ||
+ | ::<li> The band was extracted using the Sigma kit and eluted with water. | ||
+ | ::<li> Absorbance measurements indicated that no DNA was obtained, next time elution solution at 65 C will be used | ||
+ | ::<li> PCR amplification of 0A and 0B was run overnight. |
Latest revision as of 18:31, 30 July 2008
Contents |
Today in the Lab
Matt
- PCR Amplification
- Corey decided it would be best to start from the beginning and amplify the PTP2 out of Genomic DNA again. I redid the PCR and I am running it overnight.
- Inoculation
- 97 plasmid was inoculated from master plate so that it can glycerol stocked for tomorrow.
- PCR Confirmation
- Corey wanted me to do another PCR confirmation of the 97 plasmid with primers F58,F59. Expecting a band size of 2200 for this PCR when I run it on a gel for tommorrow.
Chris
- Digestion of AtCRE (again)
- AtCRE was digested with EcoRI again for one hour at 37 C
- the result was run on a 1% gel, resulting in five bands where only two were expected.
- using Vector NTI, we figured out that the incorrect enzyme was used for the digestion; in fact, EagI should have been used instead.
Dan
- PCR amplification of construct 1
- The amplification product was run on a gel however no clear bands showed up on the kodac machine. However the correct band was seen on the cutting UV machine.
- The band was extracted using the Sigma kit and eluted with water.
- Absorbance measurements indicated that no DNA was obtained, next time elution solution at 65 C will be used
- PCR amplification of 0A and 0B was run overnight.