IGEM:Harvard/2008/Lab Notebooks/DailyBook/Week10/Chemical and Light: Difference between revisions
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(New page: =Transformations in Shewie= =Getting Thermoinducible and Lac Inducible GFP into pSC101= =Getting mtrB onto thermo and lac inducible systems=) |
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=Transformations in Shewie= | =Transformations in Shewie= | ||
About 250 ng of DNA added each time. | |||
==First Set 8/22== | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Plate''' | |||
| align="center" style="background:#f0f0f0;"|'''Colonies''' | |||
| align="center" style="background:#f0f0f0;"|'''Size''' | |||
| align="center" style="background:#f0f0f0;"|'''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== | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Plate''' | |||
| align="center" style="background:#f0f0f0;"|'''Marker''' | |||
| align="center" style="background:#f0f0f0;"|'''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 Thermoinducible and Lac Inducible GFP into pSC101= | ||
=Getting mtrB onto thermo and lac inducible systems= | =Getting mtrB onto thermo and lac inducible systems= | ||
Revision as of 22:23, 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 |
