IGEM:Harvard/2008/Lab Notebooks/DailyBook/Week1/Chemical and Light: Difference between revisions
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===Overnight cultures=== | ===Overnight cultures=== | ||
06/23: The above was repeated in 5mL LB liquid cultures instead of plates. | |||
06/24: None of the cultures in LB amp grew (E1P17 in LB kan did grow). We recultured the other samples in freshly made LB amp. |
Revision as of 09:42, 24 June 2008
Chemical
Results from Friday (6/20) Electroporation and Transformation in S1
6/23:
Name Registry Name Description Origin Marker Observations P1a J23113 Low promoter GFP tester pMB1 Kan 16 pink colonies of approx 7mm diameter P2a J23150 Medium promoter GFP tester pMB1 Kan no growth P3a J23151 High promoter GFP tester pMB1 Kan 8 pink colonies of approx 7mm diameter P12 pETDuet-1 Low copy plasmid pBR322-derived ColE1 Amp Lawn P13 pCDFDuet-1 Low copy plasmid CloDF13 Sm 41 pink colonies and some small yellow ones that may be E. coli P14 pCOLADuet-1 Low copy plasmid ColA Kan No growth
Growing cultures of E. coli and S1
Picked colonies of E. coli and S1 and grew overnight at 37 and 30 degrees, respectively.
Strain Plasmid Name Registry Name Plate Information Marker Colonies Picked E. coli P15 A1 6/18 electrocompetent Cm 2 TOP10 P4 pACYC duet 6/19 restreak electrocomp Cm 2 TOP10 P7 pSB1A2 6/19 restreak electrocomp big and small Cm 2 (1 from big, 1 from small) TOP10 P8 BBa_J04450 6/19 restreak electrocomp big and small Amp 2 (1 from big, 1 from small) TOP10 P9 BBa_J04430 6/19 restreak electrocomp big and small Amp 2 (1 from big, 1 from small) TOP10 P10 BBa_I715038 6/19 restreak electrocomp big and small Amp 2 (1 from big, 1 from small) S1 P4 pACYC duet 6/18 electrocomp Cm 2 S1 P15 A1 6/19 electrocomp Cm 2 S1 P13 pCDFDuet-1 6/20 electrocomp Sm 2 S1 P1a J23113 6/20 electrocomp Kan 3 S1 P3a J23151 6/20 electrocomp Kan 3
Total number of cultures: 24
Light
Transformations of P11, P15-20
Colony counts
6/23:
Name | Description | Origin | Size | Marker | Number of Colonies |
P11 | Light responsive system, dual regulation | pUC19-derived pMB1 | 4333 | Amp/Cm | 224 |
P15 | GFP with Tet promoter | ColE1 | 937 | Amp | 360 |
P16 | Tet repressible promoter | pMB1 | 54 | Amp | 264 |
P17 | Inverter (TetR w/o promoter and Tet promoter) | OriS, P1 lytic, F1 | 902 | Kan | 2 |
P18 | lambda promoter (cI regulated) | pMB1 | 49 | Amp | 112 |
P19 | RFP with lambda promoter | pMB1 | 918 | Amp | 104 |
P20 | GFP (LVA tagged) with Lac promoter | pMB1 | 1122 | Amp, Kan | 136 |
Restreaking
6/23: P16, which has GFP under a constitutive Tet promoter did not appear to fluoresce (none of the fluorescent constructs did), so we restreaked all the of the plates (with the same antibiotic on which they were originally grown). They were left in the 37 degree incubator along with a blank Kan and a blank Amp plate (- controls).
Overnight cultures
06/23: The above was repeated in 5mL LB liquid cultures instead of plates. 06/24: None of the cultures in LB amp grew (E1P17 in LB kan did grow). We recultured the other samples in freshly made LB amp.