Team:UNICAMP-EMSE Brazil/Notebook/22 August 2011
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- | [[Image:UNICAMP-EMSE Brazil Gel 22 08.jpg|thumb|right|300px|<font color: blue>'''Electrophoresis Gel 1, | + | [[Image:UNICAMP-EMSE Brazil Gel 22 08.jpg|thumb|right|300px|<font color: blue>'''Electrophoresis Gel 1, 22/08/2011.'''</font> <br>Gel 1:Terminator/Terminator/RBS+GFP/RBS+GFP/RBS+HlyD/RBS+HlyB/RBS+TolC/HlyA/RBS+SoxR/RBS+QseB+RBS+QseC .]] |
- | == | + | ==22 August 2011== |
'''Objective:''' Cut the band of the opened vector (terminator) or cut gene (all the rest); | '''Objective:''' Cut the band of the opened vector (terminator) or cut gene (all the rest); | ||
*'''Ladder:''' 100 bp Bio-Rad | *'''Ladder:''' 100 bp Bio-Rad | ||
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:*Quantification (volume of ladder applied 6 uL; sample: 5 uL DNA +1 uL buffer ) | :*Quantification (volume of ladder applied 6 uL; sample: 5 uL DNA +1 uL buffer ) | ||
- | <br>[[Image:UNICAMP-EMSE_Brazil_Gel_22_08_2.jpg|thumb|right|300px|<font color: blue>'''Second Electrophoresis Gel, | + | <br>[[Image:UNICAMP-EMSE_Brazil_Gel_22_08_2.jpg|thumb|right|300px|<font color: blue>'''Second Electrophoresis Gel, 22/08/2011.'''</font> <br>Gel 2: Terminator/ Terminator / RBS+GFP/ RBS+GFP / RBS+HlyD/ RBS+HlyB/ RBS+TolC/ HlyA /RBS+SoxR /RBS+QseB+RBS+QseC .]] |
*'''Samples:''' Terminator/ Terminator / RBS+GFP/ RBS+GFP / RBS+HlyD/ RBS+HlyB/ RBS+TolC/ HlyA /RBS+SoxR /RBS+QseB+RBS+QseC | *'''Samples:''' Terminator/ Terminator / RBS+GFP/ RBS+GFP / RBS+HlyD/ RBS+HlyB/ RBS+TolC/ HlyA /RBS+SoxR /RBS+QseB+RBS+QseC |
Revision as of 16:23, 23 August 2011
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22 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/ 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 |
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? |
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/ HlyA /RBS+SoxR /RBS+QseB+RBS+QseC
- HlyA did not appear in the gel!! Digest and purify again !!!
Nd: non digested
D: digested
D: digested