Construction

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(September 30, 2006)
(October 1, 2006)
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     <li>Prepared o/n of DH5a and DH5a-z1 for making competent cells
     <li>Prepared o/n of DH5a and DH5a-z1 for making competent cells
     <li>Mini-prepped UT3 ABCD and P0352 (2005) BC, P0456 (2005) BC
     <li>Mini-prepped UT3 ABCD and P0352 (2005) BC, P0456 (2005) BC
-
     <li>Tested (EcoRI, PstI) length of UT3 ABCD (All wrong lengths, same as Sept ## result), P0352 (2005) BC (both have only one band, retry with higher conc. DNA), P0456 (2005) BC (correct lengths)
+
     <li>Tested (EcoRI, PstI) length of UT3 ABCD (All wrong lengths, same as Sept ## result), P0352 (2005) BC (both have only one band, retry with higher DNA conc.), P0456 (2005) BC (correct lengths)
     <li>Made Plates and transformed P0452 (2006) and I12006 (2006)
     <li>Made Plates and transformed P0452 (2006) and I12006 (2006)
     <li>Printed/posted protocols
     <li>Printed/posted protocols
Line 19: Line 19:
     <li>Finish up ligation of UT1 and P0352 and P0456 (Heat inactivate)
     <li>Finish up ligation of UT1 and P0352 and P0456 (Heat inactivate)
     <li>Transform and plate UT4 (UT1 + P0352 (2005)) ABC and UT5 (UT1 + P0456 (2005)) ABC
     <li>Transform and plate UT4 (UT1 + P0352 (2005)) ABC and UT5 (UT1 + P0456 (2005)) ABC
-
     <li>Miniprep
+
     <li>Make o/n of P0452 (2006) and I12006 (2006) for miniprep
 +
    <li>Make DH5a and DH5a-z1 competent cells
 +
    <li>Plan testing using DH5a cells
</ul>
</ul>
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 +
== September 30, 2006 ==
== September 30, 2006 ==

Revision as of 05:56, 2 October 2006

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

Contents

October 1, 2006

Charles, Andy, Anne:

  • Prepared o/n of DH5a and DH5a-z1 for making competent cells
  • Mini-prepped UT3 ABCD and P0352 (2005) BC, P0456 (2005) BC
  • Tested (EcoRI, PstI) length of UT3 ABCD (All wrong lengths, same as Sept ## result), P0352 (2005) BC (both have only one band, retry with higher DNA conc.), P0456 (2005) BC (correct lengths)
  • Made Plates and transformed P0452 (2006) and I12006 (2006)
  • Printed/posted protocols
  • Ligated UT1 ABC to P0352 A (2005) and P0456 A (2005) overnight

To-Do List:

  • Finish up ligation of UT1 and P0352 and P0456 (Heat inactivate)
  • Transform and plate UT4 (UT1 + P0352 (2005)) ABC and UT5 (UT1 + P0456 (2005)) ABC
  • Make o/n of P0452 (2006) and I12006 (2006) for miniprep
  • Make DH5a and DH5a-z1 competent cells
  • Plan testing using DH5a cells

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

September 30, 2006

Charles:

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

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 P0452 (2006) and I12006 (2006)
  • Double Digest (total volume = 30 uL) P0352 and P0456 (2005) with EcoI/XbaI in Buffer 2 and then do a gel extract/purification
  • 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

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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

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