Team:University of Ottawa/24 July 2008

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(New page: ==Today in the lab== '''Chris''' :'''Double-Check of AtCRE''' :::<li> Digested samples showing desired bands with EcoRI for one hour. :::<li> Ran against a water control and DQ2325601 dige...)
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==Today in the lab==
==Today in the lab==
'''Chris'''
'''Chris'''
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:::<li> Ran against a water control and DQ2325601 digested with EcoRI for one and a half hours on a 1% gel at 80V
:::<li> Ran against a water control and DQ2325601 digested with EcoRI for one and a half hours on a 1% gel at 80V
:::<li> One of two samples still showed desired bands following double-check. This sample was streaked onto a master plate, which was incubated at 37 C overnight, and glycerol stocked in the -80 C fridge for further use.
:::<li> One of two samples still showed desired bands following double-check. This sample was streaked onto a master plate, which was incubated at 37 C overnight, and glycerol stocked in the -80 C fridge for further use.
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'''Matt'''
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:'''Ligation Results'''
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::<li> A gel was run after two hours for ligation of PTP2/pSSA42 - unsuccessful.
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::<li> A gel was run with overnight ligation - vector and H2O controls were clean and ligation product band is slightly visible in 3:1 ratio. However there is auto ligation occuring with the vector.
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:'''PCR'''
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::<li> Dan and I decided that the initial extraction of PTP2 after PCR yielded a product that was too concentrated to be properly extracted - so I am redoing the PCR amplification with elongation time raised to 1 min 15 sec and annealing temp raised to 58 C.
 +
::<li> I will also spread the PTP2 amplification product over many different lanes on the gel so that it can be properly extracted.
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:'''Digestion'''
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::<li> pSSA42 was redigested with BamHI and xhoI.
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'''Dan'''
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:'''Wet Ware''
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::<li> Worked a little bit on trying to get some wet ware tools working.
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:'''Innoculation''
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::<li> Innoculated 6 colonies and a control

Latest revision as of 14:40, 12 August 2008

Untitled Document

 

 


Today in the lab

Chris

Double-Check of AtCRE
  • Digested samples showing desired bands with EcoRI for one hour.
  • Ran against a water control and DQ2325601 digested with EcoRI for one and a half hours on a 1% gel at 80V
  • One of two samples still showed desired bands following double-check. This sample was streaked onto a master plate, which was incubated at 37 C overnight, and glycerol stocked in the -80 C fridge for further use.

    • Matt

    Ligation Results
  • A gel was run after two hours for ligation of PTP2/pSSA42 - unsuccessful.
  • A gel was run with overnight ligation - vector and H2O controls were clean and ligation product band is slightly visible in 3:1 ratio. However there is auto ligation occuring with the vector.
    • PCR
  • Dan and I decided that the initial extraction of PTP2 after PCR yielded a product that was too concentrated to be properly extracted - so I am redoing the PCR amplification with elongation time raised to 1 min 15 sec and annealing temp raised to 58 C.
  • I will also spread the PTP2 amplification product over many different lanes on the gel so that it can be properly extracted.
    • Digestion
  • pSSA42 was redigested with BamHI and xhoI.

    • Dan

    'Wet Ware
  • Worked a little bit on trying to get some wet ware tools working.
    • 'Innoculation
  • Innoculated 6 colonies and a control
    • Retrieved from "http://2008.igem.org/Team:University_of_Ottawa/24_July_2008"