Team:UNICAMP-EMSE Brazil/Notebook/22 August 2011
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Notebook
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21 August 2011
Objective: Cut the band of the opened vector (terminator) or cut gene (all the rest);
- Ladder: 100 bp Bio-Rad
- Samples:
- (all digested, forgot to apply the closed vector sample in the gel) Terminator/Terminator/RBS+GFP/RBS+GFP/RBS+HlyD/RBS+HlyB/RBS+TolC/RBS+HlyA/RBS+SoxR/RBS+QseB+RBS+QseC
- Results:
Gene | Total size (gene+vector) | Linear vector size | Gene size | Result |
---|---|---|---|---|
Terminator | ~3300 pb | 3300 pb | No cut | Ok, complete digestion |
RBS+GFP | ~2800 pb | 2079 pb | 735 pb | Ok, complete digestion |
RBS+HlyD | 4296 pb | 2800 pb | 1496 pb | Ok, complete digestion |
RBS+HlyB | 4983 pb | 2800 pb | 2183 pb | Parcial digestion, where´s the linear vector? superposition? |
RBS+TolC | 4380 pb | 2800 pb | 1580 pb | Ok, complete digestion |
RBS+HlyA | 3027 pb | 2800 pb | 227 pb | Ok, complete digestion |
RBS+SoxR | 3323 pb | 2800 pb | 523 pb | Ok, complete digestion |
RBS+QseB+RBS+QseC | 4886 pb | 2800 pb | 2086 pb | Parcial digestion, where´s the linear vector? superposition? |
[edit]
All the genes were purified using Fermentas kit (25 uL elution) and quantified through electrophoresis:
- Ladder: 100 bp Bio-Rad
- Objective:
- Quantification (volume of ladder applied 6 uL; sample: 5 uL DNA +1 uL buffer )
- Samples: Terminator/ Terminator / RBS+GFP/ RBS+GFP / RBS+HlyD/ RBS+HlyB/ RBS+TolC/ RBS+HlyA /RBS+SoxR /RBS+QseB+RBS+QseC
- Picture legend:
- Gel: flhDC C1 (nd)/flhDC 1 (d)/flhDC 2 (nd)/ flhDC 2 (d)/ SoxS 3 (nd)/SoxS 3 (d)/SoxS 4 (nd)/ SoxS 4 (d)/ Constitutive (strong) promotor 1 (nd)/ Constitutive (strong) promotor 1 (d)/ Constitutive (strong) promotor 3 (nd)/ Constitutive (strong) promotor 3 (d)/ Constitutive (strong) promotor 3 (nd)/ Constitutive (strong) promotor 3 (d)
Nd: non digested
D: digested
D: digested