Difference between revisions of "IGEM:Harvard/2008/Lab Notebooks/DailyBook/Week10/Chemical and Light"

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(Transformations in Shewie)
(Getting mtrB onto thermo and lac inducible systems)
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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.