Team:University of Ottawa/15 July 2008

From 2008.igem.org

(Difference between revisions)
(Today on the Lab)
(Today on the Lab)
Line 17: Line 17:
:'''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.

Revision as of 19:58, 16 July 2008

Contents

Today on 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.