Team:UNICAMP-EMSE Brazil/Notebook/7 September 2011

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(Created page with "{{Template:UNICAMP-EMSE Brazil_Notebook}} <div style="text-align: left;"> [[Image:UNICAMP-EMSE Brazil Gel1 07 09 11.jpg|thumb|right|300px|<font color: blue>'''Electrophoresis Gel...")
 
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*'''Expected sizes: '''
*'''Expected sizes: '''
:* 1784bp / ~1050bp - both are OK
:* 1784bp / ~1050bp - both are OK
 +
 +
[[Image:UNICAMP-EMSE Brazil Gel2 07 09 11.jpg|thumb|right|300px|<font color: blue>'''Electrophoresis Gel 2, 07/09/2011.'''</font> <br>Gel 2: flhDC_RBS_IL12(C1) / RBS_GFP_HlyA_Term(C2) ]]
 +
 +
'''Objective:''' 
 +
:*Electrophoresis to cut and purify bands
 +
'''Samples:''' 
 +
 +
:*'''Gel 1:'''
 +
::*flhDC_RBS_IL12(C1) / RBS_GFP_HlyA_Term(C2)
 +
 +
*'''Ladder:'''
 +
:*100-10.000 bp (Fermentas)
 +
*'''Gel Agarose concentration:'''
 +
:*1,0%
 +
 +
===Digestions recipes for gel purification:===
 +
*'''RBS+GFP+HlyA+T (X-P)'''
 +
:*32ul  - RBS+GFP+HlyA+T miniprep (C2)
 +
:*4ul    - 10X Tango Buffer
 +
:*1.4ul  - XbaI
 +
:*2.6ul  - PstI
 +
:*'''TOTAL = 40ul'''
 +
 +
*'''flhDC+RBS+IL-12 (E-S)'''
 +
:*31ul  - flhDC+RBS+IL-12 miniprep (did on 06/09/2011)
 +
:*4ul  - 10X Buffer R
 +
:*1ul  - EcoRI
 +
:*4ul  - SpeI
 +
:*'''TOTAL = 40ul'''
 +
 +
*'''flhDC (S-P) '''
 +
:*32ul  - flhDC miniprep
 +
:*4ul    - 10X Tango Buffer
 +
:*1.4ul  - SpeI
 +
:*2.6ul  - PstI
 +
:*'''TOTAL = 40ul'''
 +
 +
*'''HlyA+T (E-X)'''
 +
:*29ul  - HlyA+T miniprep
 +
:*8ul    - 10X Tango Buffer
 +
:*1ul    - EcoRI
 +
:*2ul  - XbaI
 +
:*'''TOTAL = 40ul'''
 +
 +
'''Tasks:'''<br>
 +
Incubate reactions at 37°C for 4hours.<br>
 +
Electrophoresis (loading the whole digestions volume - join two wells in the gel)<br>
 +
Purify the bands<br>
 +
Electrophoresis to confirm (load 5ul of gel purified digestion)<br>

Latest revision as of 01:41, 29 September 2011

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Notebook

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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]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/3_October_2011&action=edit 3] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/4_October_2011&action=edit 4] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/5_October_2011&action=edit 5] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/6_October_2011&action=edit 6] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/7_October_2011&action=edit 7] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/8_October_2011&action=edit 8] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/9_October_2011&action=edit 9]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/10_October_2011&action=edit 10] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/11_October_2011&action=edit 11] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/12_October_2011&action=edit 12] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/13_October_2011&action=edit 13] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/14_October_2011&action=edit 14] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/15_October_2011&action=edit 15] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/16_October_2011&action=edit 16]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/17_October_2011&action=edit 17] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/18_October_2011&action=edit 18] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/19_October_2011&action=edit 19] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/20_October_2011&action=edit 20] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/21_October_2011&action=edit 21] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/22_October_2011&action=edit 22] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/23_October_2011&action=edit 23]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/24_October_2011&action=edit 24] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/25_October_2011&action=edit 25] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/26_October_2011&action=edit 26] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/27_October_2011&action=edit 27] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/28_October_2011&action=edit 28] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/29_October_2011&action=edit 29] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/30_October_2011&action=edit 30]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/31_October_2011&action=edit 31]


Electrophoresis Gel 1, 07/09/2011.
Gel 1: flhDC_RBS_IL12(C1)(E - S) / RBS_GFP_HlyA_Term(C2)(X - P)

07 September 2011

Objective:

  • Electrophoresis to cut and purify bands

Samples:

  • Gel 1:
  • flhDC_RBS_IL12(C1)(E - S) / RBS_GFP_HlyA_Term(C2)(X - P)
  • Ladder:
  • 100-10.000 bp (Fermentas)
  • Gel Agarose concentration:
  • 1,0%
  • Expected sizes:
  • 1784bp / ~1050bp - both are OK
Electrophoresis Gel 2, 07/09/2011.
Gel 2: flhDC_RBS_IL12(C1) / RBS_GFP_HlyA_Term(C2)

Objective:

  • Electrophoresis to cut and purify bands

Samples:

  • Gel 1:
  • flhDC_RBS_IL12(C1) / RBS_GFP_HlyA_Term(C2)
  • Ladder:
  • 100-10.000 bp (Fermentas)
  • Gel Agarose concentration:
  • 1,0%

Digestions recipes for gel purification:

  • RBS+GFP+HlyA+T (X-P)
  • 32ul - RBS+GFP+HlyA+T miniprep (C2)
  • 4ul - 10X Tango Buffer
  • 1.4ul - XbaI
  • 2.6ul - PstI
  • TOTAL = 40ul
  • flhDC+RBS+IL-12 (E-S)
  • 31ul - flhDC+RBS+IL-12 miniprep (did on 06/09/2011)
  • 4ul - 10X Buffer R
  • 1ul - EcoRI
  • 4ul - SpeI
  • TOTAL = 40ul
  • flhDC (S-P)
  • 32ul - flhDC miniprep
  • 4ul - 10X Tango Buffer
  • 1.4ul - SpeI
  • 2.6ul - PstI
  • TOTAL = 40ul
  • HlyA+T (E-X)
  • 29ul - HlyA+T miniprep
  • 8ul - 10X Tango Buffer
  • 1ul - EcoRI
  • 2ul - XbaI
  • TOTAL = 40ul

Tasks:
Incubate reactions at 37°C for 4hours.
Electrophoresis (loading the whole digestions volume - join two wells in the gel)
Purify the bands

Electrophoresis to confirm (load 5ul of gel purified digestion)
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