Team:University of Ottawa/15 July 2008

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Today in the Lab
Tammy
 * Transformation of XL10 Competent E. coli cells with pDR197:AtCKX2
 * <li>1 uL Pure pDR197:AtCKX2
 * <li>1 ul 1:10 dilution pDR197:AtCKX2
 * <li>1 ul 1:100 dilution pDR197:AtCKX2
 * <li>100 ul XL10 E.Coli per reaction
 * <li>900 ul LB Broth per reaction
 * <li>3 LB Ampicillin (50 ug/mL) Plates
 * <li>Began Incubation of transformed cells at 1:50 pm.

Matt
 * Inoculation of 97 - BY4742 cells
 * <li> I inoculated the 97 BY4742 cells once again for glycerol stock tomorrow
 * PCR confirmation
 * <li> 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
 * <li> 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
 * <li> AtCRE was digested using EagI at 37 C for about one hour
 * <li> the result was run on a 1% gel, which showed two bands, both of which were expected
 * Gel Extraction of AtCRE
 * <li> the AtCRE was extracted from the gel using the gel extraction kit and protocol
 * <li> unfortunately, the final AtCRE sample was vacuumed up with the other waste by accident
 * 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.

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.