IGEM:Harvard/2008/Lab Notebooks/DailyBook/Week2/Chemical and Light: Difference between revisions
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##Add promoters. | ##Add promoters. | ||
#Make primers that have BioBricks prefix and suffix to flank the origin on Duet Vectors so we can take it out. | #Make primers that have BioBricks prefix and suffix to flank the origin on Duet Vectors so we can take it out. | ||
#PCR out mtrA and mtrB from Shewie. Also put in with repressors. | |||
==Re-Transformation of BioBrick Parts in E1 and E3== | ==Re-Transformation of BioBrick Parts in E1 and E3== | ||
Revision as of 08:39, 30 June 2008
Chemical
Goals for Chemical Group
- Transform LacI, TetR, cI + their promoters and GFP. Refer [here].
- Cut Lac promoter -> GFP (P20), and Tet promoter -> GFP (P15), with Xba and PstI and cut SB3K3 (P5) with Xba and PstI and ligate.
- Transform cI promoter. (Refer [here].
- Put constitutive promoters on TetR, LacI, cI.
- Put terminators and RBS on TetR, LacI, and cI.
- Add promoters.
- Make primers that have BioBricks prefix and suffix to flank the origin on Duet Vectors so we can take it out.
- PCR out mtrA and mtrB from Shewie. Also put in with repressors.
Re-Transformation of BioBrick Parts in E1 and E3
Name Registry Name Description Origin Marker Transformed? (6/26) Picked colonies? (6/29) Miniprepped? Made glycerol stocks? P7 pSB1A3* High copy plasmid with Amp resistance ColE1 Amp Failed in E1 P8 BBa_J04450 RFP with LacI promoter pMB1 Amp Failed in E1 P9 BBa_J04430 GFP with LacI promoter pMB1 Amp Failed in E1 P10 BBa_I715038 T7 Polymerase with LacI promoter pMB1 Kan/Amp Failed in E1 P36 BBa_I763004 GFP + IPTG + LVA Kan/Amp Failed in E1 P5 (2007) pSB3K3* Low-Medium copy vector (w/ death gene) p15A Kan Yes, in E3 2 P6 (2007) BBa-E1010 RFP only (w/ death gene) pMB1 Kan Failed in E3 P5 (2008) pSB3K3* Low-Medium copy vector (w/ death gene) p15A Kan Failed in E3
Name Registry Name Description Origin Marker Transformed? (6/27) Picked colonies? (6/29) Miniprepped? Made glycerol stocks? P12 pETDuet-1 Low copy plasmid pBR322-derived ColE1 Amp in E1 2 P37 BBa_I722007 Constructive expression LacI w/ pTetR promoter Amp in E1 2 P38 BBa_J23114 High constitutive promoter Amp in E1 2 P38 (2007) BBa_J23114 High constitutive promoter Amp in E1 2 P39 BBa_J23113 Low constitutive promoter Amp in E1 2 P39 (2007) BBa_J23113 Low constitutive promoter Amp in E1 2 P40 BBa_B0032 Ribosome Binding Site Amp in E1 2 P40 (2007) BBa_B0032 Ribosome Binding Site Amp in E1 2 P41 BBa_B0010 Transcriptional Terminator Amp in E1 2 P41 (2007) BBa_B0010 Transcriptional Terminator Amp in E1 P42 BBa_C0012 LacI coding region Amp in E1 2 P42 (2007) BBa_C0012 LacI coding region Amp in E1 1 P43 [1] TetR coding region Amp in E1 2 P43 (2007) [2] TetR coding region Amp in E1 2 P44 BBa_C0051 cI lambda Amp in E1 2 P44 (2007) BBa_C0051 cI lambda Amp in E1 2 P45 BBa_E0240 GFP only w/ RBS & terminator Amp in E1 2 P45 (2007) BBa_E0240 GFP only w/ RBS & terminator Amp in E1 2 P46 BBa_I51020 Base Vector Amp in E3 2 P47 BBa_P1003 Kan Resistance Cassette Kan No in E1 0 P47 (2007) BBa_P1003 Kan Resistance Cassette Kan in E1 2 P48 BBa_P1004 Cm Resistance Cassette Cm No in E1 0 P48 (2007) BBa_P1004 Cm Resistance Cassette Cm in E1 2 P49 BBa_P1002 Amp Resistance Cassette Amp in E1 P49 (2007) BBa_P1002 Amp Resistance Cassette Amp in E1 2 P50 BBa_P1005 Tet Resistance Cassette Amp in E1 P50 (2007) BBa_P1005 Tet Resistance Cassette Amp in E1 2
