Team:UNICAMP-EMSE Brazil/Notebook/9 September 2011
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[[Image:UNICAMP-EMSE Brazil Gel1 09 09 11.jpg|thumb|right|300px|<font color: blue>'''Electrophoresis Gel 1, 09/09/2011.'''</font> <br>Gel 1: SoxS+RBS+GFP+HlyA+T nd/ SoxS+RBS+GFP+HlyA+T E-P/ Const. Promoter+ RBS+SoxR+ T nd/ Const. Promoter+ RBS+SoxR+ T E-P/ SoxS+RBS+HlyB+HlyD+TolC + T nd/ SoxS+RBS+HlyB+HlyD+TolC + T E-P ]] | [[Image:UNICAMP-EMSE Brazil Gel1 09 09 11.jpg|thumb|right|300px|<font color: blue>'''Electrophoresis Gel 1, 09/09/2011.'''</font> <br>Gel 1: SoxS+RBS+GFP+HlyA+T nd/ SoxS+RBS+GFP+HlyA+T E-P/ Const. Promoter+ RBS+SoxR+ T nd/ Const. Promoter+ RBS+SoxR+ T E-P/ SoxS+RBS+HlyB+HlyD+TolC + T nd/ SoxS+RBS+HlyB+HlyD+TolC + T E-P ]] | ||
- | + | [[Image:UNICAMP-EMSE Brazil Gel2 09 09 11.jpg|thumb|right|300px|<font color: blue>'''Electrophoresis Gel 2, 09/09/2011.'''</font> <br>Gel 2: SoxS+RBS+GFP+HlyA+T E-X/ / Const. Promoter+ RBS+SoxR+ T E-S ]] | |
+ | ==09 September 2011== | ||
+ | ===Digestion and Electrophoresis=== | ||
*Digestion recipe: | *Digestion recipe: | ||
:*DNA – 7ul | :*DNA – 7ul | ||
Line 58: | Line 60: | ||
::*SoxS+RBS+GFP+HlyA+T E-X/ / Const. Promoter+ RBS+SoxR+ T E-S | ::*SoxS+RBS+GFP+HlyA+T E-X/ / Const. Promoter+ RBS+SoxR+ T E-S | ||
- | [[Image:UNICAMP-EMSE Brazil | + | |
+ | ===Check up electrophoresis=== | ||
+ | [[Image:UNICAMP-EMSE Brazil Gel3 09 09 11.jpg|thumb|right|300px|<font color: blue>'''Electrophoresis Gel 3, 09/09/2011.'''</font> <br>Gel 2: Sox S + RBS + IL10 ; flHDC + RBS +IL12+ HlyA + T (digest by (E-P))]] | ||
+ | '''Objective:''' | ||
+ | :*Checking digestion | ||
+ | |||
+ | '''Samples:''' | ||
+ | :*'''Gel 3:''' | ||
+ | ::*Sox S + RBS + IL10 ; flHDC + RBS +IL12+ HlyA + T (digest by (E-P)) | ||
+ | |||
+ | *'''Ladder:''' | ||
+ | :*100-10.000 bp (Fermentas) | ||
+ | *'''Gel Agarose concentration:''' | ||
+ | :*1,0% | ||
+ | |||
+ | <br clear="all" /> | ||
+ | *'''<font size=+5 color=teal>Results:''' | ||
+ | </font> | ||
+ | <font align=center> | ||
+ | |||
+ | {|border="1" cellpadding="5" cellspacing="0" align="center" | ||
+ | |- | ||
+ | ! scope="col" style="background:#efefef;" | Construction | ||
+ | ! scope="col" style="background:#efefef;" | Total size (gene+vector) (bp) | ||
+ | ! scope="col" style="background:#efefef;" | Linear vector size (bp) | ||
+ | ! scope="col" style="background:#efefef;" | Gene size (bp) | ||
+ | ! scope="col" style="background:#efefef;" | Result | ||
+ | |- | ||
+ | | Sox S + RBS + IL10 | ||
+ | | 3543 | ||
+ | | 2800 | ||
+ | | 743 | ||
+ | | No Gene size band | ||
+ | |||
+ | |- | ||
+ | | Sox S + RBS + IL10 | ||
+ | | 3543 | ||
+ | | 2800 | ||
+ | | 743 | ||
+ | | No Gene size band | ||
+ | |||
+ | |- | ||
+ | | Sox S + RBS + IL10 | ||
+ | | 3543 | ||
+ | | 2800 | ||
+ | | 743 | ||
+ | | No Gene size band; and partial digestion | ||
+ | |||
+ | |- | ||
+ | | flHDC +RBS +IL12+ HlyA +T | ||
+ | | 5311 | ||
+ | | 3205 | ||
+ | | 2106 | ||
+ | | Ok | ||
+ | |||
+ | |- | ||
+ | | flHDC +RBS +IL12+ HlyA +T | ||
+ | | 5311 | ||
+ | | 3205 | ||
+ | | 2106 | ||
+ | | Ok | ||
+ | |- | ||
+ | |}</font> |
Latest revision as of 21:23, 28 September 2011
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09 September 2011
Digestion and Electrophoresis
- Digestion recipe:
- DNA – 7ul
- Buffer O – 2ul
- EcoRI – 0,5ul
- PstI – 0,5ul
- Water – 10ul
Samples:
- Gel 1:
- SoxS+RBS+GFP+HlyA+T nd/ SoxS+RBS+GFP+HlyA+T E-P/ Const. Promoter+ RBS+SoxR+ T nd/ Const. Promoter+ RBS+SoxR+ T E-P/ SoxS+RBS+HlyB+HlyD+TolC + T nd/ SoxS+RBS+HlyB+HlyD+TolC + T E-P
- Ladder:
- 100-10.000 bp (Fermentas)
- Gel Agarose concentration:
- 1,0%
- Results:
Construction | Total size (gene+vector) (bp) | Linear vector size (bp) | Gene size (bp) | Result |
---|---|---|---|---|
1) SoxS+RBS+GFP+HlyA+T | 4365 | 3205 | 1160 | ok |
2) Const. Promoter+ RBS+SoxR+ T | 2732 | 2079 | 653 | ok |
3) SoxS+RBS+HlyB+HlyD+TolC + T | 8257 | 2800 | 5457 | ?? |
Samples:
- Gel 2:
- SoxS+RBS+GFP+HlyA+T E-X/ / Const. Promoter+ RBS+SoxR+ T E-S
Check up electrophoresis
Objective:
- Checking digestion
Samples:
- Gel 3:
- Sox S + RBS + IL10 ; flHDC + RBS +IL12+ HlyA + T (digest by (E-P))
- Ladder:
- 100-10.000 bp (Fermentas)
- Gel Agarose concentration:
- 1,0%
- Results:
Construction | Total size (gene+vector) (bp) | Linear vector size (bp) | Gene size (bp) | Result |
---|---|---|---|---|
Sox S + RBS + IL10 | 3543 | 2800 | 743 | No Gene size band |
Sox S + RBS + IL10 | 3543 | 2800 | 743 | No Gene size band |
Sox S + RBS + IL10 | 3543 | 2800 | 743 | No Gene size band; and partial digestion |
flHDC +RBS +IL12+ HlyA +T | 5311 | 3205 | 2106 | Ok |
flHDC +RBS +IL12+ HlyA +T | 5311 | 3205 | 2106 | Ok |