Construction
From 2006.igem.org
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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]
