Construction

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

September 29, 2006

Charles:

  • Prepared o/n of competent cells (DH5a-z1 and DH5a), and UT3 ABCD to miniprep again

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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