Team:UNICAMP-EMSE Brazil/Notebook/17 September 2011
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[[Image:UNICAMP-EMSE Brazil Gel4 17 09 11.jpg|thumb|right|300px|<font color: blue>'''Electrophoresis Gel 4, 17/09/2011.'''</font> <br>Gel 4: Ladder / pConst+RBS+SoxR+T C1 (D)/ pConst+RBS+SoxR+T C2 (D)/ pConst+RBS+SoxR+T C3 (D)/ pConst+RBS+SoxR+T C4 (D)/ pConst+RBS+SoxR+T (ND) / SoxS+RBS+IL-10+HlyA+T C1 (D)/ SoxS+RBS+IL-10+HlyA+T C2 (D)/ SoxS+RBS+IL-10+HlyA+T C3 (D)/ SoxS+RBS+IL-10+HlyA+T C4 (D)/ SoxS+RBS+IL-10+HlyA+T (ND)/ flhDC+RBS+IL-12+HlyA+T (D) / flhDC+RBS+IL-12+HlyA+T (ND)/ flhDC+RBS+GFP+HlyA+T (D)/ flhDC+RBS+GFP+HlyA+T (ND)/ SoxS+RBS+GFP+HlyA+T (D)/ SoxS+RBS+GFP+HlyA+T Test (D)/ SoxS+RBS+GFP+HlyA+T (ND) / pConst+RBS+QseB+RBS+QseC+T (D)/ pConst+RBS+QseB+RBS+QseC+T (ND) ]] | [[Image:UNICAMP-EMSE Brazil Gel4 17 09 11.jpg|thumb|right|300px|<font color: blue>'''Electrophoresis Gel 4, 17/09/2011.'''</font> <br>Gel 4: Ladder / pConst+RBS+SoxR+T C1 (D)/ pConst+RBS+SoxR+T C2 (D)/ pConst+RBS+SoxR+T C3 (D)/ pConst+RBS+SoxR+T C4 (D)/ pConst+RBS+SoxR+T (ND) / SoxS+RBS+IL-10+HlyA+T C1 (D)/ SoxS+RBS+IL-10+HlyA+T C2 (D)/ SoxS+RBS+IL-10+HlyA+T C3 (D)/ SoxS+RBS+IL-10+HlyA+T C4 (D)/ SoxS+RBS+IL-10+HlyA+T (ND)/ flhDC+RBS+IL-12+HlyA+T (D) / flhDC+RBS+IL-12+HlyA+T (ND)/ flhDC+RBS+GFP+HlyA+T (D)/ flhDC+RBS+GFP+HlyA+T (ND)/ SoxS+RBS+GFP+HlyA+T (D)/ SoxS+RBS+GFP+HlyA+T Test (D)/ SoxS+RBS+GFP+HlyA+T (ND) / pConst+RBS+QseB+RBS+QseC+T (D)/ pConst+RBS+QseB+RBS+QseC+T (ND) ]] | ||
[[Image:UNICAMP-EMSE Brazil Gel5 17 09 11.jpg|thumb|right|300px|<font color: blue>'''Electrophoresis Gel 5, 17/09/2011.'''</font> <br>Gel 5: Ladder / flhDC+RBS+HlyB+RBS+HlyD+TolC+T (D)/ flhDC+RBS+HlyB+RBS+HlyD+TolC+T (ND)/ SoxS+RBS+ HlyB+RBS+HlyD+TolC+T (D)/ SoxS+RBS+ HlyB+RBS+HlyD+TolC+T (ND)/ pSB1C3 TolC C10 (D) / pSB1C3 TolC C10 (ND) ]] | [[Image:UNICAMP-EMSE Brazil Gel5 17 09 11.jpg|thumb|right|300px|<font color: blue>'''Electrophoresis Gel 5, 17/09/2011.'''</font> <br>Gel 5: Ladder / flhDC+RBS+HlyB+RBS+HlyD+TolC+T (D)/ flhDC+RBS+HlyB+RBS+HlyD+TolC+T (ND)/ SoxS+RBS+ HlyB+RBS+HlyD+TolC+T (D)/ SoxS+RBS+ HlyB+RBS+HlyD+TolC+T (ND)/ pSB1C3 TolC C10 (D) / pSB1C3 TolC C10 (ND) ]] | ||
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*'''<font size=+5 color=teal>Results:''' | *'''<font size=+5 color=teal>Results:''' | ||
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+ | {|border="1" cellpadding="5" cellspacing="0" align="center" | ||
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+ | ! scope="col" style="background:#efefef;" | Construction | ||
+ | ! scope="col" style="background:#efefef;" | Total size (gene+vector) | ||
+ | ! scope="col" style="background:#efefef;" | Linear vector size | ||
+ | ! scope="col" style="background:#efefef;" | Gene size | ||
+ | ! scope="col" style="background:#efefef;" | Result | ||
+ | |}</font> | ||
+ | <br> | ||
+ | <br> | ||
<html> | <html> |
Revision as of 01:54, 18 September 2011
Home | Project | Methods | Results | Data | Team | Notebook | Human Practices | Safety | Profile | Sponsors | Wix |
Contents |
Notebook
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17 September 2011
Sequences to ship
Quantification and checking digestion of sequences to ship
Objectives:
- Quantification of linearized pSB1C3 vector purified from the gel (volume of ladder applied 6 uL; sample: 5 uL DNA +1 uL buffer),
- Checking digestion ( EcoRI and PstI) of samples mini-preped 09/15 and 09/16.
- Checking digestion of SoxS+HlyB+HlyD +TolC+T C1 1/ SoxS+HlyB+HlyD +TolC+T C1 2/ SoxS+HlyB+HlyD +TolC+T C1 3 (EcoRI and PstI) of samples mini-preped 09/08.
Samples:
- Gel 1:
- pSB1/ pSB 2/ pSB 3/ pSB 4/ pSB 5/ RBS+IL-12 C4/ RBS+IL-12 C5/ RBS+HlyD C4/ RBS+HlyD C5/ RBS+Tol C C10/ RBS+Tol C C11/ RBS+QseB+RBS+QseC C10/ RBS+QseB+RBS+QseC C11/ RBS+QseB+RBS+QseC C12/
- Gel 2:
- RBS+SoxS C10/ RBS+SoxS C11/ RBS+SoxS C12/ RBS+SoxS C13/ RBS+HlyB C10/ RBS+HlyB C11/ RBS+HlyB C12/ SoxS+ RBS+HlyB +RBS+HlyD+ RBS+TolC+T C1 1/ SoxS+ RBS+HlyB+ RBS+HlyD + RBS+TolC+T C1 2/ SoxS+ RBS+HlyB+ RBS+HlyD + RBS+TolC+T C3/
- Ladder:
- 100-10.000 bp (Fermentas)
- Gel Agarose concentration:
- 1,0%
- Results:
Revising constructs
Objective:
- Checking digestion with Eco RI of some constructs that had strange result in former electrophoresis
Samples:
- Gel 3:
- (d)RBS+HlyD C4/ (nd)RBS+HlyD C4/ (d)RBS+Tol C C11/ (nd)RBS+Tol C C11/ (d)RBS+QseB+RBS+QseC C10/ (nd)RBS+QseB+RBS+QseC C10/(d)RBS+SoxS C13/ (nd)RBS+SoxS C13/ (d)RBS+HlyB C10/ (nd)RBS+HlyB C10/ (d)RBS+HlyB C11/(nd) RBS+HlyB C11/ (d)SoxS+ RBS+HlyB +RBS+HlyD+ RBS+TolC+T C1 1/(nd) SoxS+ RBS+HlyB +RBS+HlyD+ RBS+TolC+T C1 1
- Results:
Building the devices
Quantification and checking digestion of sequences to ship
Objectives:
- Finishing the assembly of our devices
- Secretion device (containing HlyB, HlyD and TolC in two manners: regulated by flhDC promoter or SoxS promoter)
- Th1-directed device (containing QseC and QseB sensor proteins constitutively expressed and IL-12 whose expression is regulated by flhDC promoter)
- Th2-directed device (containing the SoxR sensor protein constitutively expressed and IL-10 whose expression is regulated by SoxS promoter)
- After finishing the assembly of these parts, we aim to put together the plasmid containing the Secretion system (1.1) and the plasmid containing the sensor-response systems (the one that senses Adrenaline and responds by producing IL-12 (1.2) and the one which senses Nitric Oxide and responds by producing IL-10 (1.3).
Samples:
- Gel 1:
- Ladder / pConst+RBS+SoxR+T C1 (D)/ pConst+RBS+SoxR+T C2 (D)/ pConst+RBS+SoxR+T C3 (D)/ pConst+RBS+SoxR+T C4 (D)/ pConst+RBS+SoxR+T (ND) / SoxS+RBS+IL-10+HlyA+T C1 (D)/ SoxS+RBS+IL-10+HlyA+T C2 (D)/ SoxS+RBS+IL-10+HlyA+T C3 (D)/ SoxS+RBS+IL-10+HlyA+T C4 (D)/ SoxS+RBS+IL-10+HlyA+T (ND)/ flhDC+RBS+IL-12+HlyA+T (D) / flhDC+RBS+IL-12+HlyA+T (ND)/ flhDC+RBS+GFP+HlyA+T (D)/ flhDC+RBS+GFP+HlyA+T (ND)/ SoxS+RBS+GFP+HlyA+T (D)/ SoxS+RBS+GFP+HlyA+T Test (D)/ SoxS+RBS+GFP+HlyA+T (ND) / pConst+RBS+QseB+RBS+QseC+T (D)/ pConst+RBS+QseB+RBS+QseC+T (ND)
- Gel 2:
- Ladder / flhDC+RBS+HlyB+RBS+HlyD+TolC+T (D)/ flhDC+RBS+HlyB+RBS+HlyD+TolC+T (ND)/ SoxS+RBS+ HlyB+RBS+HlyD+TolC+T (D)/ SoxS+RBS+ HlyB+RBS+HlyD+TolC+T (ND)/ pSB1C3 TolC C10 (D) / pSB1C3 TolC C10 (ND)
- Ladder:
- 100-10.000 bp (Fermentas)
- Gel Agarose concentration:
- 1,0%
- Results:
Construction | Total size (gene+vector) | Linear vector size | Gene size | Result |
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