Team:University of Ottawa/15 July 2008

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__TOC__
__TOC__
-
==Today on the Lab==
+
==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'''
:<li>1 uL Pure pDR197:AtCKX2
:<li>1 uL Pure pDR197:AtCKX2
:<li>1 ul 1:10 dilution pDR197:AtCKX2
:<li>1 ul 1:10 dilution pDR197:AtCKX2
Line 13: Line 86:
:'''Inoculation of 97 - BY4742 cells'''
:'''Inoculation of 97 - BY4742 cells'''
:<li> I inoculated the 97 BY4742 cells once again for glycerol stock tomorrow
:<li> I inoculated the 97 BY4742 cells once again for glycerol stock tomorrow
-
'''PCR confirmation'''
+
:'''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.
:<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'''
+
:'''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.
:<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.

Latest revision as of 18:31, 30 July 2008

Untitled Document

 

 


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.
    • Retrieved from "http://2008.igem.org/Team:University_of_Ottawa/15_July_2008"