Team:UNICAMP-EMSE Brazil/Notebook/31 August 2011

From 2011.igem.org

(Difference between revisions)
(→Digestion, purification and ligation!!!)
 
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*'''<font size=+5 color=teal>Results:'''</font>
*'''<font size=+5 color=teal>Results:'''</font>
<font align=center>
<font align=center>
-
[[Image:UNICAMP-EMSE_Brazil_Table_temp.jpg|650px|center]]
+
{|border="1" cellpadding="5" cellspacing="0" align="center"
 +
|-
 +
! 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
 +
|-  
 +
| HlyA+Terminator 
 +
| 3527
 +
| 3205
 +
| No cut
 +
| Ok
 +
 +
|-
 +
| HlyA+Terminator 
 +
| 3527
 +
| 3205
 +
| No cut
 +
| Ok
 +
 +
|-
 +
| HlyA+Terminator 
 +
| 3527
 +
| 3205
 +
| No cut
 +
| Ok
 +
 +
|-
 +
| HlyA+Terminator 
 +
| 3527
 +
| 3205
 +
| No cut
 +
| Ok
 +
 +
|-
 +
| SoxS+RBS+GFP 
 +
| 3535
 +
| 2800
 +
| 838
 +
| Ok
 +
 +
|-
 +
| SoxS+RBS+GFP 
 +
| 3535
 +
| 2800
 +
| 838
 +
| Ok
 +
 +
|-
 +
| RBS+HlyD+TolC+Terminator 
 +
| 6376
 +
| 3205
 +
| No cut
 +
| Run more
 +
 +
|-
 +
| RBS+HlyD+TolC+Terminator 
 +
| 6376
 +
| 3205
 +
| No cut
 +
| Run more
 +
 +
|-
 +
| RBS+HlyD+TolC+Terminator 
 +
| 6376
 +
| 3205
 +
| No cut
 +
| Run more
 +
 +
|-
 +
| RBS+HlyD+TolC+Terminator 
 +
| 6376
 +
| 3205
 +
| No cut
 +
| Run more
 +
 +
|-
 +
| SoxS+RBS+IL 10 
 +
| 3543
 +
| 2800
 +
| 743
 +
| Ok
 +
 +
|-
 +
| SoxS+RBS+IL 10 
 +
| 3543
 +
| 2800
 +
| 743
 +
| Ok
 +
 +
|-
 +
| flHDC+RBS+HlyB 
 +
| 5109
 +
| 2800
 +
| 2309
 +
| Run more
 +
 +
|-
 +
| flHDC+RBS+HlyB 
 +
| 5109
 +
| 2800
 +
| 2309
 +
| Run more
 +
 +
|-
 +
| flHDC+RBS+IL 12
 +
| 4584
 +
| 2800
 +
| 1784
 +
| Where is the vector?
 +
 +
|-
 +
| flHDC+RBS+GFP 
 +
| 3661
 +
| 2800
 +
| 861
 +
| No digestion
 +
 +
|-
 +
|}</font>
<br>
<br>
::::<font size=+4 color=Purple align=center>'''OBS:'''</font><font size=+5 color=red> flHDC+RBS+IL 12 and  flHDC+RBS+GFP did not work! Inoculate again from a permanent culture.!</font>
::::<font size=+4 color=Purple align=center>'''OBS:'''</font><font size=+5 color=red> flHDC+RBS+IL 12 and  flHDC+RBS+GFP did not work! Inoculate again from a permanent culture.!</font>
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<br>
<br>
[[Image:UNICAMP-EMSE Brazil Gel2 31 08 11.jpg|thumb|right|300px|<font color: blue>'''Electrophoresis Gel 2, 31/08/2011.'''</font> <br>Gel 2: HlyA+Terminator (E-X)/ HlyA+Terminator (E-X)/ HlyA+Terminator (E-X)/ HlyA+Terminator (E-X)/  SoxS+RBS+GFP (E-S)/ SoxS+RBS+GFP (E-S)/ RBS+HlyD+TolC+Terminator (E-X)/ RBS+HlyD+TolC+Terminator (E-X)/ RBS+HlyD+TolC+Terminator (E-X)/ RBS+HlyD+TolC+Terminator (E-X)/ SoxS+RBS+IL 10 (E-S)/ SoxS+RBS+IL 10 (E-S)/ flHDC+RBS+HlyB (E-S)/ flHDC+RBS+HlyB (E-S)/flHDC+RBS+IL 12 (E-S)/ flHDC+RBS+GFP (E-S)/]]
[[Image:UNICAMP-EMSE Brazil Gel2 31 08 11.jpg|thumb|right|300px|<font color: blue>'''Electrophoresis Gel 2, 31/08/2011.'''</font> <br>Gel 2: HlyA+Terminator (E-X)/ HlyA+Terminator (E-X)/ HlyA+Terminator (E-X)/ HlyA+Terminator (E-X)/  SoxS+RBS+GFP (E-S)/ SoxS+RBS+GFP (E-S)/ RBS+HlyD+TolC+Terminator (E-X)/ RBS+HlyD+TolC+Terminator (E-X)/ RBS+HlyD+TolC+Terminator (E-X)/ RBS+HlyD+TolC+Terminator (E-X)/ SoxS+RBS+IL 10 (E-S)/ SoxS+RBS+IL 10 (E-S)/ flHDC+RBS+HlyB (E-S)/ flHDC+RBS+HlyB (E-S)/flHDC+RBS+IL 12 (E-S)/ flHDC+RBS+GFP (E-S)/]]
 +
===Same electrophoresys but after a longer period run:===
===Same electrophoresys but after a longer period run:===
*'''Ladder:'''  
*'''Ladder:'''  

Latest revision as of 12:53, 18 September 2011

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Contents

Notebook

Click on a date to see what we have done!

May
MTWTFSS
            [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/1_May_2011&action=edit 1]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/2_May_2011&action=edit 2] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/3_May_2011&action=edit 3] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/4_May_2011&action=edit 4] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/5_May_2011&action=edit 5] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/6_May_2011&action=edit 6] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/7_May_2011&action=edit 7] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/8_May_2011&action=edit 8]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/9_May_2011&action=edit 9] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/10_May_2011&action=edit 10] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/11_May_2011&action=edit 11] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/12_May_2011&action=edit 12] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/13_May_2011&action=edit 13] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/14_May_2011&action=edit 14] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/15_May_2011&action=edit 15]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/16_May_2011&action=edit 16] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/17_May_2011&action=edit 17] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/18_May_2011&action=edit 18] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/19_May_2011 19] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/20_May_2011&action=edit 20] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/21_May_2011&action=edit 21] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/22_May_2011&action=edit 22]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/23_May_2011&action=edit 23] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/24_May_2011&action=edit 24] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/25_May_2011 25] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/26_May_2011 26] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/27_May_2011 27] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/28_May_2011&action=edit 28] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/29_May_2011&action=edit 29]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/30_May_2011 30] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/31_May_2011 31]
June
MTWTFSS
    [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/1_June_2011 1] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/2_June_2011 2] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/3_June_2011 3] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/4_June_2011&action=edit 4] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/5_June_2011&action=edit 5]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/6_June_2011 6] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/7_June_2011 7] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/8_June_2011 8] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/9_June_2011 9] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/10_June_2011 10] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/11_June_2011&action=edit 11] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/12_June_2011&action=edit 12]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/13_June_2011 13] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/14_June_2011&action=edit 14] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/15_June_2011 15] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/16_June_2011 16] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/17_June_2011 17] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/18_June_2011&action=edit 18] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/19_June_2011&action=edit 19]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/20_June_2011 20] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/21_June_2011 21] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/22_June_2011 22] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/23_June_2011&action=edit 23] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/24_June_2011&action=edit 24] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/25_June_2011&action=edit 25] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/26_June_2011&action=edit 26]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/27_June_2011&action=edit 27] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/28_June_2011&action=edit 28] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/29_June_2011 29] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/30_June_2011&action=edit 30]
July
MTWTFSS
        [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/1_July_2011&action=edit 1] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/2_July_2011&action=edit 2] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/3_July_2011 3]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/4_July_2011&action=edit 4] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/5_July_2011&action=edit 5] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/6_July_2011&action=edit 6] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/7_July_2011 7] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/8_July_2011 8] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/9_July_2011&action=edit 9] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/10_July_2011&action=edit 10]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/11_July_2011 11] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/12_July_2011 12] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/13_July_2011 13] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/14_July_2011 14] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/15_July_2011 15] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/16_July_2011 16] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/17_July_2011&action=edit 17]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/18_July_2011 18] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/19_July_2011 19] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/20_July_2011&action=edit 20] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/21_July_2011 21] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/22_July_2011 22] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/23_July_2011&action=edit 23] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/24_July_2011&action=edit 24]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/25_July_2011 25] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/26_July_2011 26] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/27_July_2011 27] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/28_July_2011 28] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/29_July_2011&action=edit 29] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/30_July_2011&action=edit 30] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/31_July_2011&action=edit 31]
August
MTWTFSS
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/1_August_2011 1] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/2_August_2011 2] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/3_August_2011&action=edit 3] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/4_August_2011&action=edit 4] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/5_August_2011&action=edit 5] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/6_August_2011&action=edit 6] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/7_August_2011&action=edit 7]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/8_August_2011&action=edit 8] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/9_August_2011&action=edit 9] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/10_August_2011&action=edit 10] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/11_August_2011&action=edit 11] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/12_August_2011&action=edit 12] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/13_August_2011&action=edit 13] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/14_August_2011&action=edit 14]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/15_August_2011&action=edit 15] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/16_August_2011&action=edit 16] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/17_August_2011&action=edit 17] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/18_August_2011&action=edit 18] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/19_August_2011 19] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/20_August_2011 20] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/21_August_2011 21]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/22_August_2011 22] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/23_August_2011 23] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/24_August_2011 24] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/25_August_2011 25] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/26_August_2011 26] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/27_August_2011 27] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/28_August_2011&action=edit 28]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/29_August_2011 29] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/30_August_2011 30] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/31_August_2011 31]
September
MTWTFSS
      [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/1_September_2011 1] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/2_September_2011 2] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/3_September_2011&action=edit 3] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/4_September_2011&action=edit 4]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/5_September_2011 5] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/6_September_2011 6] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/7_September_2011 7] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/8_September_2011 8] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/9_September_2011 9] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/10_September_2011 10] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/11_September_2011 11]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/12_September_2011 12] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/13_September_2011 13] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/14_September_2011 14] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/15_September_2011&action=edit 15] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/16_September_2011 16] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/17_September_2011 17] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/18_September_2011&action=edit 18]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/19_September_2011 19] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/20_September_2011&action=edit 20] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/21_September_2011 21] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/22_September_2011 22] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/23_September_2011 23] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/24_September_2011 24] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/25_September_2011 25]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/26_September_2011 26] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/27_September_2011 27] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/28_September_2011 28] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/29_September_2011 29] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/30_September_2011&action=edit 30]
October
MTWTFSS
          [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/1_October_2011&action=edit 1] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/2_October_2011 2]
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[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/31_October_2011&action=edit 31]


Electrophoresis Gel 1, 31/08/2011.
Gel 1: HlyA+Terminator (E-X)/ HlyA+Terminator (E-X)/ HlyA+Terminator (E-X)/ HlyA+Terminator (E-X)/ SoxS+RBS+GFP (E-S)/ SoxS+RBS+GFP (E-S)/ RBS+HlyD+TolC+Terminator (E-X)/ RBS+HlyD+TolC+Terminator (E-X)/ RBS+HlyD+TolC+Terminator (E-X)/ RBS+HlyD+TolC+Terminator (E-X)/ SoxS+RBS+IL 10 (E-S)/ SoxS+RBS+IL 10 (E-S)/ flHDC+RBS+HlyB (E-S)/ flHDC+RBS+HlyB (E-S)/flHDC+RBS+IL 12 (E-S)/ flHDC+RBS+GFP (E-S)/

31 August 2011

Digestion, purification and ligation!!!

Objective:

  • Digestion of constructs to purify and ligate them.

Samples:

  • Gel 1:
  • HlyA+Terminator (E-X)/ HlyA+Terminator (E-X)/ HlyA+Terminator (E-X)/ HlyA+Terminator (E-X)/ SoxS+RBS+GFP (E-S)/ SoxS+RBS+GFP (E-S)/ RBS+HlyD+TolC+Terminator (E-X)/ RBS+HlyD+TolC+Terminator (E-X)/ RBS+HlyD+TolC+Terminator (E-X)/ RBS+HlyD+TolC+Terminator (E-X)/ SoxS+RBS+IL 10 (E-S)/ SoxS+RBS+IL 10 (E-S)/ flHDC+RBS+HlyB (E-S)/ flHDC+RBS+HlyB (E-S)/flHDC+RBS+IL 12 (E-S)/ flHDC+RBS+GFP (E-S)/
  • Ladder:
  • 100-10.000 bp (Fermentas)
  • Gel Agarose concentration:
  • 1,0%


  • Results:

Construction Total size (gene+vector) Linear vector size Gene size Result
HlyA+Terminator 3527 3205 No cut Ok
HlyA+Terminator 3527 3205 No cut Ok
HlyA+Terminator 3527 3205 No cut Ok
HlyA+Terminator 3527 3205 No cut Ok
SoxS+RBS+GFP 3535 2800 838 Ok
SoxS+RBS+GFP 3535 2800 838 Ok
RBS+HlyD+TolC+Terminator 6376 3205 No cut Run more
RBS+HlyD+TolC+Terminator 6376 3205 No cut Run more
RBS+HlyD+TolC+Terminator 6376 3205 No cut Run more
RBS+HlyD+TolC+Terminator 6376 3205 No cut Run more
SoxS+RBS+IL 10 3543 2800 743 Ok
SoxS+RBS+IL 10 3543 2800 743 Ok
flHDC+RBS+HlyB 5109 2800 2309 Run more
flHDC+RBS+HlyB 5109 2800 2309 Run more
flHDC+RBS+IL 12 4584 2800 1784 Where is the vector?
flHDC+RBS+GFP 3661 2800 861 No digestion


OBS: flHDC+RBS+IL 12 and flHDC+RBS+GFP did not work! Inoculate again from a permanent culture.!





Electrophoresis Gel 2, 31/08/2011.
Gel 2: HlyA+Terminator (E-X)/ HlyA+Terminator (E-X)/ HlyA+Terminator (E-X)/ HlyA+Terminator (E-X)/ SoxS+RBS+GFP (E-S)/ SoxS+RBS+GFP (E-S)/ RBS+HlyD+TolC+Terminator (E-X)/ RBS+HlyD+TolC+Terminator (E-X)/ RBS+HlyD+TolC+Terminator (E-X)/ RBS+HlyD+TolC+Terminator (E-X)/ SoxS+RBS+IL 10 (E-S)/ SoxS+RBS+IL 10 (E-S)/ flHDC+RBS+HlyB (E-S)/ flHDC+RBS+HlyB (E-S)/flHDC+RBS+IL 12 (E-S)/ flHDC+RBS+GFP (E-S)/

Same electrophoresys but after a longer period run:

  • Ladder:
  • 100-10.000 bp (Fermentas)
  • Gel Agarose concentration:
  • 1,0%

Samples:

  • Gel 2: Same of Gel1!
  • HlyA+Terminator (E-X)/ HlyA+Terminator (E-X)/ HlyA+Terminator (E-X)/ HlyA+Terminator (E-X)/ SoxS+RBS+GFP (E-S)/ SoxS+RBS+GFP (E-S)/ RBS+HlyD+TolC+Terminator (E-X)/ RBS+HlyD+TolC+Terminator (E-X)/ RBS+HlyD+TolC+Terminator (E-X)/ RBS+HlyD+TolC+Terminator (E-X)/ SoxS+RBS+IL 10 (E-S)/ SoxS+RBS+IL 10 (E-S)/ flHDC+RBS+HlyB (E-S)/ flHDC+RBS+HlyB (E-S)/flHDC+RBS+IL 12 (E-S)/ flHDC+RBS+GFP (E-S)/

Results:

  • RBS+HlyD+TolC+Terminator has two bands, but the smaller one can be the closed vector while the larger ones can be the linearized vector. We cut the larger ones.


Quantification of digestion:

Objective:

Electrophoresis Gel 3, 31/08/2011.
Gel 3: HlyA+T (E-X) 1/ HlyA+T (E-X) 2/ HlyA+T (E-X) 3/ HlyA+T (E-X) 4/ SoxS+RBS+IL10 (E-S) 1/ SoxS+RBS+IL10 (E-S) 2/ RBS+HlyD+TolC+T (E-X) 1/ RBS+HlyD+TolC+T (E-X) 2/ RBS+HlyD+TolC+T (E-X) 3/ RBS+HlyD+TolC+T (E-X) 4/ / flHDC+RBS+HlyB (E-S) 1/ flHDC+RBS+HlyB (E-S) 2/ SoxS+RBS+GFP (E-S) 1/ SoxS+RBS+GFP (E-S) 1
  • Quantification of samples purified from the gel
  • Ladder:
  • 100-10.000 bp (Fermentas)
  • Gel Agarose concentration:
  • 1,0%
  • Gel 3:
  • HlyA+T (E-X) 1/ HlyA+T (E-X) 2/ HlyA+T (E-X) 3/ HlyA+T (E-X) 4/ SoxS+RBS+IL10 (E-S) 1/ SoxS+RBS+IL10 (E-S) 2/ RBS+HlyD+TolC+T (E-X) 1/ RBS+HlyD+TolC+T (E-X) 2/ RBS+HlyD+TolC+T (E-X) 3/ RBS+HlyD+TolC+T (E-X) 4/ / flHDC+RBS+HlyB (E-S) 1/ flHDC+RBS+HlyB (E-S) 2/ SoxS+RBS+GFP (E-S) 1/ SoxS+RBS+GFP (E-S) 1



  • Other tasks made today:
  1. Ligation of samples purified from the gel
  2. Transformation and platting
  3. Inoculation of fLHDC+RBS+GFP C6 from permanent culture and new colonies of fLHDC+RBS+ IL12 from the plate (C8, C9,C10,C11).





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