Team:Groningen/project notebook/10 August 2011

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(Difference between revisions)
(Created page with "{{HeaderGroningen2011}} {{notebookgroningen}} '''Joyce''' <br> <br> colony PCR of PBAD-pSB1C3: <br> Composition master mix: <br> 10× Taq buffer: 40μl <br> dNTPs 10mM: 8μl <br...")
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{{HeaderGroningen2011}} {{notebookgroningen}}  
{{HeaderGroningen2011}} {{notebookgroningen}}  
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=Wed 10 August 2011=
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'''Vessa'''
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==To Do List==
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*Transformation---2nd batch (ON ligation mixture)
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*Screening transforman ---Growth ON culture:
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**12 variant autoinducing construts
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**RFP internal control --> PCR colonies screening
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**construct 21.12 (pSB1K3 revDT+P(LasR)+LasR-TAG) 
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'''Implementation'''
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RBS-RFP PCR colonies screening --> pick 9 single clones
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<br> 15.5 ul MQ
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<br> 2 ul buffer
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<br> 1.2 ul MgCl2
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<br> 0.4 ul dNTPs
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<br> 0.4 ul Forward
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<br> 0.4 ul Reverse
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<br> 0.1 ul Taq DNA pol
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<br> DNA sample
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Growth ON culture for plasmid isolation from
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<br> 7 plates 22.xx : 2 clones each
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<br> 1 plate 21.12 : 2 clones
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'''Results'''
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None of growing clones have RBS-RFP insert --> self-ligation/incomplete vector digestion
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'''Joyce'''
'''Joyce'''
<br>
<br>

Revision as of 11:53, 21 September 2011


March
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[http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/7_March_2011&preload=Team:Groningen/project_notebook/Day&action=edit 7] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/8_March_2011&preload=Team:Groningen/project_notebook/Day&action=edit 8] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/9_March_2011&preload=Team:Groningen/project_notebook/Day&action=edit 9] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/10_March_2011&preload=Team:Groningen/project_notebook/Day&action=edit 10] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/11_March_2011&preload=Team:Groningen/project_notebook/Day&action=edit 11] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/12_March_2011&preload=Team:Groningen/project_notebook/Day&action=edit 12] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/13_March_2011&preload=Team:Groningen/project_notebook/Day&action=edit 13]
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[http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/28_March_2011&preload=Team:Groningen/project_notebook/Day&action=edit 28] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/29_March_2011&preload=Team:Groningen/project_notebook/Day&action=edit 29] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/30_March_2011&preload=Team:Groningen/project_notebook/Day&action=edit 30] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/31_March_2011&preload=Team:Groningen/project_notebook/Day&action=edit 31]
April
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[http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/11_April_2011&preload=Team:Groningen/project_notebook/Day&action=edit 11] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/12_April_2011&preload=Team:Groningen/project_notebook/Day&action=edit 12] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/13_April_2011&preload=Team:Groningen/project_notebook/Day&action=edit 13] [http://2011.igem.org/Team:Groningen/project_notebook/14_April_2011 14] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/15_April_2011&preload=Team:Groningen/project_notebook/Day&action=edit 15] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/16_April_2011&preload=Team:Groningen/project_notebook/Day&action=edit 16] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/17_April_2011&preload=Team:Groningen/project_notebook/Day&action=edit 17]
[http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/18_April_2011&preload=Team:Groningen/project_notebook/Day&action=edit 18] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/19_April_2011&preload=Team:Groningen/project_notebook/Day&action=edit 19] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/20_April_2011&preload=Team:Groningen/project_notebook/Day&action=edit 20] [http://2011.igem.org/Team:Groningen/project_notebook/21_April_2011 21] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/22_April_2011&preload=Team:Groningen/project_notebook/Day&action=edit 22] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/23_April_2011&preload=Team:Groningen/project_notebook/Day&action=edit 23] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/24_April_2011&preload=Team:Groningen/project_notebook/Day&action=edit 24]
[http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/25_April_2011&preload=Team:Groningen/project_notebook/Day&action=edit 25] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/26_April_2011&preload=Team:Groningen/project_notebook/Day&action=edit 26] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/27_April_2011&preload=Team:Groningen/project_notebook/Day&action=edit 27] [http://2011.igem.org/Team:Groningen/project_notebook/28_April_2011 28] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/29_April_2011&preload=Team:Groningen/project_notebook/Day&action=edit 29] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/30_April_2011&preload=Team:Groningen/project_notebook/Day&action=edit 30]
May
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[http://2011.igem.org/Team:Groningen/project_notebook/2_May_2011 2] [http://2011.igem.org/Team:Groningen/project_notebook/3_May_2011 3] [http://2011.igem.org/Team:Groningen/project_notebook/4_May_2011 4] [http://2011.igem.org/Team:Groningen/project_notebook/5_May_2011 5] [http://2011.igem.org/Team:Groningen/project_notebook/6_May_2011 6] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/7_May_2011&preload=Team:Groningen/project_notebook/Day&action=edit 7] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/8_May_2011&preload=Team:Groningen/project_notebook/Day&action=edit 8]
[http://2011.igem.org/Team:Groningen/project_notebook/9_May_2011 9] [http://2011.igem.org/Team:Groningen/project_notebook/10_May_2011 10] [http://2011.igem.org/Team:Groningen/project_notebook/11_May_2011 11] [http://2011.igem.org/Team:Groningen/project_notebook/12_May_2011 12] [http://2011.igem.org/Team:Groningen/project_notebook/13_May_2011 13] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/14_May_2011&preload=Team:Groningen/project_notebook/Day&action=edit 14] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/15_May_2011&preload=Team:Groningen/project_notebook/Day&action=edit 15]
[http://2011.igem.org/Team:Groningen/project_notebook/16_May_2011 16] [http://2011.igem.org/Team:Groningen/project_notebook/17_May_2011 17] [http://2011.igem.org/Team:Groningen/project_notebook/18_May_2011 18] [http://2011.igem.org/Team:Groningen/project_notebook/19_May_2011 19] [http://2011.igem.org/Team:Groningen/project_notebook/20_May_2011 20] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/21_May_2011&preload=Team:Groningen/project_notebook/Day&action=edit 21] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/22_May_2011&preload=Team:Groningen/project_notebook/Day&action=edit 22]
[http://2011.igem.org/Team:Groningen/project_notebook/23_May_2011 23] [http://2011.igem.org/Team:Groningen/project_notebook/24_May_2011 24] [http://2011.igem.org/Team:Groningen/project_notebook/25_May_2011 25] [http://2011.igem.org/Team:Groningen/project_notebook/26_May_2011 26] [http://2011.igem.org/Team:Groningen/project_notebook/27_May_2011 27] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/28_May_2011&preload=Team:Groningen/project_notebook/Day&action=edit 28] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/29_May_2011&preload=Team:Groningen/project_notebook/Day&action=edit 29]
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June
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[http://2011.igem.org/Team:Groningen/project_notebook/6_June_2011 6] [http://2011.igem.org/Team:Groningen/project_notebook/7_June_2011 7] [http://2011.igem.org/Team:Groningen/project_notebook/8_June_2011 8] [http://2011.igem.org/Team:Groningen/project_notebook/9_June_2011 9] [http://2011.igem.org/Team:Groningen/project_notebook/10_June_2011 10] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/11_June_2011&preload=Team:Groningen/project_notebook/Day&action=edit 11] [http://2011.igem.org/Team:Groningen/project_notebook/12_June_2011 12]
[http://2011.igem.org/Team:Groningen/project_notebook/13_June_2011 13] [http://2011.igem.org/Team:Groningen/project_notebook/14_June_2011 14] [http://2011.igem.org/Team:Groningen/project_notebook/15_June_2011 15] [http://2011.igem.org/Team:Groningen/project_notebook/16_June_2011 16] [http://2011.igem.org/Team:Groningen/project_notebook/17_June_2011 17] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/18_June_2011&preload=Team:Groningen/project_notebook/Day&action=edit 18] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/19_June_2011&preload=Team:Groningen/project_notebook/Day&action=edit 19]
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[http://2011.igem.org/Team:Groningen/project_notebook/27_June_2011 27] [http://2011.igem.org/Team:Groningen/project_notebook/28_June_2011 28] [http://2011.igem.org/Team:Groningen/project_notebook/29_June_2011 29] [http://2011.igem.org/Team:Groningen/project_notebook/30_June_2011 30]
July
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[http://2011.igem.org/Team:Groningen/project_notebook/4_July_2011 4] [http://2011.igem.org/Team:Groningen/project_notebook/5_July_2011 5] [http://2011.igem.org/Team:Groningen/project_notebook/6_July_2011 6] [http://2011.igem.org/Team:Groningen/project_notebook/7_July_2011 7] [http://2011.igem.org/Team:Groningen/project_notebook/8_July_2011 8] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/9_July_2011&preload=Team:Groningen/project_notebook/Day&action=edit 9] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/10_July_2011&preload=Team:Groningen/project_notebook/Day&action=edit 10]
[http://2011.igem.org/Team:Groningen/project_notebook/11_July_2011 11] [http://2011.igem.org/Team:Groningen/project_notebook/12_July_2011 12] [http://2011.igem.org/Team:Groningen/project_notebook/13_July_2011 13] [http://2011.igem.org/Team:Groningen/project_notebook/14_July_2011 14] [http://2011.igem.org/Team:Groningen/project_notebook/15_July_2011 15] [http://2011.igem.org/Team:Groningen/project_notebook/16_July_2011 16] [http://2011.igem.org/Team:Groningen/project_notebook/17_July_2011 17]
[http://2011.igem.org/Team:Groningen/project_notebook/18_July_2011 18] [http://2011.igem.org/Team:Groningen/project_notebook/19_July_2011 19] [http://2011.igem.org/Team:Groningen/project_notebook/20_July_2011 20] [http://2011.igem.org/Team:Groningen/project_notebook/21_July_2011 21] [http://2011.igem.org/Team:Groningen/project_notebook/22_July_2011 22] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/23_July_2011&preload=Team:Groningen/project_notebook/Day&action=edit 23] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/24_July_2011&preload=Team:Groningen/project_notebook/Day&action=edit 24]
[http://2011.igem.org/Team:Groningen/project_notebook/25_July_2011 25] [http://2011.igem.org/Team:Groningen/project_notebook/26_July_2011 26] [http://2011.igem.org/Team:Groningen/project_notebook/27_July_2011 27] [http://2011.igem.org/Team:Groningen/project_notebook/28_July_2011 28] [http://2011.igem.org/Team:Groningen/project_notebook/29_July_2011 29] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/30_July_2011&preload=Team:Groningen/project_notebook/Day&action=edit 30] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/31_July_2011&preload=Team:Groningen/project_notebook/Day&action=edit 31]
August
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[http://2011.igem.org/Team:Groningen/project_notebook/1_August_2011 1] [http://2011.igem.org/Team:Groningen/project_notebook/2_August_2011 2] [http://2011.igem.org/Team:Groningen/project_notebook/3_August_2011 3] [http://2011.igem.org/Team:Groningen/project_notebook/4_August_2011 4] [http://2011.igem.org/Team:Groningen/project_notebook/5_August_2011 5] [http://2011.igem.org/Team:Groningen/project_notebook/6_August_2011 6] [http://2011.igem.org/Team:Groningen/project_notebook/7_August_2011 7]
[http://2011.igem.org/Team:Groningen/project_notebook/8_August_2011 8] [http://2011.igem.org/Team:Groningen/project_notebook/9_August_2011 9] [http://2011.igem.org/Team:Groningen/project_notebook/10_August_2011 10] [http://2011.igem.org/Team:Groningen/project_notebook/11_August_2011 11] [http://2011.igem.org/Team:Groningen/project_notebook/12_August_2011 12] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/13_August_2011&preload=Team:Groningen/project_notebook/Day&action=edit 13] [http://2011.igem.org/Team:Groningen/project_notebook/14_August_2011 14]
[http://2011.igem.org/Team:Groningen/project_notebook/15_August_2011 15] [http://2011.igem.org/Team:Groningen/project_notebook/16_August_2011 16] [http://2011.igem.org/Team:Groningen/project_notebook/17_August_2011 17] [http://2011.igem.org/Team:Groningen/project_notebook/18_August_2011 18] [http://2011.igem.org/Team:Groningen/project_notebook/19_August_2011 19] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/20_August_2011&preload=Team:Groningen/project_notebook/Day&action=edit 20] [http://2011.igem.org/Team:Groningen/project_notebook/21_August_2011 21]
[http://2011.igem.org/Team:Groningen/project_notebook/22_August_2011 22] [http://2011.igem.org/Team:Groningen/project_notebook/23_August_2011 23] [http://2011.igem.org/Team:Groningen/project_notebook/24_August_2011 24] [http://2011.igem.org/Team:Groningen/project_notebook/25_August_2011 25] [http://2011.igem.org/Team:Groningen/project_notebook/26_August_2011 26] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/27_August_2011&preload=Team:Groningen/project_notebook/Day&action=edit 27] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/28_August_2011&preload=Team:Groningen/project_notebook/Day&action=edit 28]
[http://2011.igem.org/Team:Groningen/project_notebook/29_August_2011 29] [http://2011.igem.org/Team:Groningen/project_notebook/30_August_2011 30] [http://2011.igem.org/Team:Groningen/project_notebook/31_August_2011 31]
September
MTWTFSS
      [http://2011.igem.org/Team:Groningen/project_notebook/1_September_2011 1] [http://2011.igem.org/Team:Groningen/project_notebook/2_September_2011 2] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/3_September_2011&preload=Team:Groningen/project_notebook/Day&action=edit 3] [http://2011.igem.org/Team:Groningen/project_notebook/4_September_2011 4]
[http://2011.igem.org/Team:Groningen/project_notebook/5_September_2011 5] [http://2011.igem.org/Team:Groningen/project_notebook/6_September_2011 6] [http://2011.igem.org/Team:Groningen/project_notebook/7_September_2011 7] [http://2011.igem.org/Team:Groningen/project_notebook/8_September_2011 8] [http://2011.igem.org/Team:Groningen/project_notebook/9_September_2011 9] [http://2011.igem.org/Team:Groningen/project_notebook/10_September_2011 10] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/11_September_2011&preload=Team:Groningen/project_notebook/Day&action=edit 11]
[http://2011.igem.org/Team:Groningen/project_notebook/12_September_2011 12] [http://2011.igem.org/Team:Groningen/project_notebook/13_September_2011 13] [http://2011.igem.org/Team:Groningen/project_notebook/14_September_2011 14] [http://2011.igem.org/Team:Groningen/project_notebook/15_September_2011 15] [http://2011.igem.org/Team:Groningen/project_notebook/16_September_2011 16] [http://2011.igem.org/Team:Groningen/project_notebook/17_September_2011 17] [http://2011.igem.org/Team:Groningen/project_notebook/18_September_2011 18]
[http://2011.igem.org/Team:Groningen/project_notebook/19_September_2011 19] [http://2011.igem.org/Team:Groningen/project_notebook/20_September_2011 20] [http://2011.igem.org/Team:Groningen/project_notebook/21_September_2011 21] [http://2011.igem.org/Team:Groningen/project_notebook/22_September_2011 22] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/23_September_2011&preload=Team:Groningen/project_notebook/Day&action=edit 23] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/24_September_2011&preload=Team:Groningen/project_notebook/Day&action=edit 24] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/25_September_2011&preload=Team:Groningen/project_notebook/Day&action=edit 25]
[http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/26_September_2011&preload=Team:Groningen/project_notebook/Day&action=edit 26] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/27_September_2011&preload=Team:Groningen/project_notebook/Day&action=edit 27] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/28_September_2011&preload=Team:Groningen/project_notebook/Day&action=edit 28] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/29_September_2011&preload=Team:Groningen/project_notebook/Day&action=edit 29] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/30_September_2011&preload=Team:Groningen/project_notebook/Day&action=edit 30]
October
MTWTFSS
          [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/1_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 1] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/2_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 2]
[http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/3_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 3] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/4_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 4] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/5_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 5] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/6_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 6] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/7_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 7] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/8_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 8] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/9_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 9]
[http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/10_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 10] [http://2011.igem.org/Team:Groningen/project_notebook/11_October_2011 11] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/12_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 12] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/13_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 13] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/14_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 14] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/15_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 15] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/16_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 16]
[http://2011.igem.org/Team:Groningen/project_notebook/17_October_2011 17] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/18_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 18] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/19_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 19] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/20_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 20] [http://2011.igem.org/Team:Groningen/project_notebook/21_October_2011 21] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/22_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 22] [http://2011.igem.org/Team:Groningen/project_notebook/23_October_2011 23]
[http://2011.igem.org/Team:Groningen/project_notebook/24_October_2011 24] [http://2011.igem.org/Team:Groningen/project_notebook/25_October_2011 25] [http://2011.igem.org/Team:Groningen/project_notebook/26_October_2011 26] [http://2011.igem.org/Team:Groningen/project_notebook/27_October_2011 27] [http://2011.igem.org/Team:Groningen/project_notebook/28_October_2011 28] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/29_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 29] [http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/30_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 30]
[http://2011.igem.org/wiki/index.php?title=Team:Groningen/project_notebook/31_October_2011&preload=Team:Groningen/project_notebook/Day&action=edit 31]

Wed 10 August 2011

Vessa

To Do List

  • Transformation---2nd batch (ON ligation mixture)
  • Screening transforman ---Growth ON culture:
    • 12 variant autoinducing construts
    • RFP internal control --> PCR colonies screening
    • construct 21.12 (pSB1K3 revDT+P(LasR)+LasR-TAG)

Implementation RBS-RFP PCR colonies screening --> pick 9 single clones
15.5 ul MQ
2 ul buffer
1.2 ul MgCl2
0.4 ul dNTPs
0.4 ul Forward
0.4 ul Reverse
0.1 ul Taq DNA pol
DNA sample

Growth ON culture for plasmid isolation from
7 plates 22.xx : 2 clones each
1 plate 21.12 : 2 clones

Results None of growing clones have RBS-RFP insert --> self-ligation/incomplete vector digestion

Joyce

colony PCR of PBAD-pSB1C3:
Composition master mix:
10× Taq buffer: 40μl
dNTPs 10mM: 8μl
MgCl2: 24μl
Forward biobrickvector primer: 8μl
Reverse biobrickvector primer: 8μl
Taq polymerase: 2μl
MQ water: 310μl

PCR conditions:
Pre heated lid: 111°C
Denaturation: 94°C for 10min
Cycle (33×)
Denaturation: 94°C for 30s
Annealing: 60°C for 30s
Extenstion: 72°C for 30s
Final extension: 72°C for 10min
Store infinite at 4°C
Analyse PCR samples on a 1% agarosegel

Plasmid prep of overnight culture PBAD-RBS-GFP colonies 5 and 6
Measure DNA concentration: 40 ng/microliter
Samples were send for sequencing and glycerol stocks were made

dsDNA arrived!!:)
structure in plasmid: SacI-prefix-taRNA-suffix-BamHI-prefix-crRNA-suffix-SalI-prefix-PRM-suffix-HindIII.
Dissolve the dsDNA in 50 microliter of MQ water, pipet 10 times up and down and incubate it for one hour at room temperature
Than make a dilution 100× and use 1 microliter for transformation:
Transformation dsDNA
Add to 40μl competent cells 10μl ligation mixture
Incubate for 30 min on ice
Heat shock: 45s at 42 degrees
Incubate cells on ice for 2 min
Add 1ml LB medium+ 25mM glucose
Incubate cells for 1h/1.5h at 37 degrees
Spin the cells down, resuspend them in 100μl LB medium and plate 90μl and 10μl out on the plates
Put the plates in the stove at 37 degrees overnight

Make overnight culture of the PBAD-pSB1C3 colonies 1 and 2