Construction

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<font size=4>< [[Aug11-Sep7]] | [[Sep8-14]] | [[Sep15-26]] | [[Construction]]></font>
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<font size=4>< [[Aug11-Sep7]] | [[Sep8-14]] | [[Sep15-26]] | [[Construction]] ></font>
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== September 30, 2006 ==
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'''Charles:'''
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<ul>
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    <li>Prepare o/n of UT3 ABCD and P0352, P0456 (2005) (2 vials each)
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    <li>Print/posted protocols
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</ul>
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'''To-Do List:'''
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<ul>
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    <li>Prepare o/n of DH5a and DH5a-z1 for making competent cells
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    <li>Miniprep UT3 ABCD and P0352, P0456 (2005) and test
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    <li>Make Plates and transform P0
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    <li>Double Digest (volume X3) P0352 and P0456 (2005) with EcoI/XbaI in Buffer 2 and extract
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    <li>Ligate UT1 and P0352/P0456 (3 tries each)
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</ul>
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[http://2006.igem.org/University_of_Toronto_2006 Home]
== September 29, 2006 ==
== September 29, 2006 ==

Revision as of 21:50, 30 September 2006

< Aug11-Sep7 | Sep8-14 | Sep15-26 | Construction >

Contents

September 30, 2006

Charles:

  • Prepare o/n of UT3 ABCD and P0352, P0456 (2005) (2 vials each)
  • Print/posted protocols

To-Do List:

  • Prepare o/n of DH5a and DH5a-z1 for making competent cells
  • Miniprep UT3 ABCD and P0352, P0456 (2005) and test
  • Make Plates and transform P0
  • Double Digest (volume X3) P0352 and P0456 (2005) with EcoI/XbaI in Buffer 2 and extract
  • Ligate UT1 and P0352/P0456 (3 tries each)

[http://2006.igem.org/University_of_Toronto_2006 Home]

September 29, 2006

Charles:

  • Prepared o/n of DH5a-z1 and UT3 ABCD to miniprep again
  • Plated DH5a

To-Do List:

  • Make new competent DH5a-z1
  • Prepare o/n of regular DH5a
  • Plated P0 (2006) using new competent cells
  • Double digest UT1 (host) with SpeI/PstI in Buffer 2 and double digest P0352/P0456 (2005) with XbaI/PstI in Buffer 3

[http://2006.igem.org/University_of_Toronto_2006 Home]

September 28, 2006

Andy:

  • Miniprep P0456 (2005)
  • Checked the lengths (EcoRI and PstI, Buffer EcoRI): UT3 ABCDE, P0456 (2005) and P0352 (2005)
  • Transformed P0152, P0352, P0452, and P0456 (2006). [Plated on Amp plates, because no more Amp/Kan plate]

To-Do List:

  • Plate DH5a cells
  • Make LB Broth, plates and print protocols
  • Digest P0352 and P0456 (2005) and ligate with UT1
  • Prepare o/n of UT3 ABCDE again and try to miniprep again

[http://2006.igem.org/University_of_Toronto_2006 Home]

September 27, 2006

Charles:

  • Made freezer stocks of UT2 x 2 and UT3 x 2

Andy, Natalie:

  • Mini-preppred UT3 B(AB) C(BB) D(BC) E(CC)
  • Transformed and plated P0452 (2005) (On Amp rather than proper AK)
  • Checked lengths of UT3 BCDE (inconclusive due to anomalous bands, D appears promising - yellow rating), P0152 (2nd check, Extra band, wrong insert length), P0352 (2nd check, not the best bands, correct lengths - changed rating from red to yellow)
  • Prepared o/n of P0456 (2005) for Miniprep
  • Added part lengths and resistance info in spreadsheet

To-Do List:

  • Miniprep P0456 (2005)
  • Take P0456, P0452, P0152, P0352 from 2006 registry
  • Check lengths of UT3 BCDE and P0352 again
  • Make more plates, make more LB
  • Learn to use new DH5a (with no LacI expression) for testing
  • Post up new protocols

[http://2006.igem.org/University_of_Toronto_2006 Home]

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