IGEM:Harvard/2008/Lab Notebooks/DailyBook/Week10/Chemical and Light: Difference between revisions
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=Getting mtrB onto thermo and lac inducible systems= | =Getting mtrB onto thermo and lac inducible systems= | ||
==Ligating mtrB TOPO with RBS== | |||
Grew up more mtrB TOPO for next week. | |||
==PCR of mtrB with RBS + mtrB primers== | |||
Scheduled for next week. |
Revision as of 22:25, 30 August 2008
Transformations in Shewie
About 250 ng of DNA added each time.
First Set 8/22
Plate | Colonies | Size | Color |
S1 P102 (P108+45) Kan | 5 colonies | 3mm to 5 mm | Pink |
S1 P121+P38 (P124) 1:2.5 Kan A | 1 colony | 5mm to 7.5mm | Pink |
S1 P28 (ColE1) Kan | 0 colonies | ||
S1 P121+P38(P124) 1:2.5 Kan B | 0 colonies | ||
S1 P121+P39 1:2.5 Dephos Kan A | 0 colonies | ||
S1 Topo vector (puc) 2ul Kan X-gal | >50 colonies | 1mm | White |
S1 Topo vector (puc) 4ul Kan X-gal | >50 colonies | 1mm | White |
S1 P121+P39 1:2.5 Kan D | 0 colonies | ||
S1 (-) ctrl (just cells) Kan | 0 colonies | ||
S1 (+) ctrl (E1 p59b) Kan | 2 colonies | 5mm | Pink |
Poor transformation efficiency in the positive control suggests that there may be a problem with our cells. All samples, except for P102, were retransformed the following week with fresh cell cultures.
Second Set (with new cells) 8/27
Plate | Marker | Description |
S1 P124 1:2.5 A | Kan | 14 2-3mm pink colonies, ~100 0.5mm white colonies |
S1 P124 1:2.5 B | Kan | 2 pink 2mm colonies, >100 0.5mm white colonies |
S1 P125 1:2.5 Dephos A | Kan | 2 pink 2mm colonies, >100 0.5mm white colonies |
S1 P125 1:2.5 D | Kan | 8 2mm pink colonies, ~100 0.5mm white colonies |
S1 P28 | Kan | ~8 1-2mm pink colonies, ~50 0.5mm white colonies |
S1 TOPO 2uL | Kan | Lawn of (>200) 0.5mm white colonies |
S1 TOPO 4 uL (w/ Xgal) | Kan | Lawn of (>200) 0.5mm white colonies |
S1 P59b (+) ctrl | Kan | ~50 2mm pink colonies, ~50 0.5mm white colonies |
S1 (-) ctrl | Kan | 50-100 0.5mm white colonies |
Getting Thermoinducible and Lac Inducible GFP into pSC101
Getting mtrB onto thermo and lac inducible systems
Ligating mtrB TOPO with RBS
Grew up more mtrB TOPO for next week.
PCR of mtrB with RBS + mtrB primers
Scheduled for next week.