Team:Hawaii/Notebook/2008-09-23

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(Difference between revisions)
(margaret)
(MCS replacment redo. PCR amplify BB insert for replacing pRL1383a-BamHI-HindIII with BB sites)
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:*PCR of rep, ran a gel to verify size, and exo-sap. will send in tomorrow
:*PCR of rep, ran a gel to verify size, and exo-sap. will send in tomorrow
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===PCR: pRL1383a MCS Replacement===
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:'''Norman'''
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* x6 parallel MCS replacements fragment by PCR amplification.  PCR out each MCS replacement insert with HindIII-VF2BB_fx._sb.1 and BamHI-VRBB_rx._sb.1
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*# BBa_I52002 from pSB4A5 (Spring 2008 Plate 1022 1C)
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*# BBa_I52001 from pSB4T5 (Spring 2008 Plate 1020 1A)
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*# BBa_I52002 from BBa_I51020 (BBa_I51020 Base Vector from glycerol stock Inventory [http://packrat.stjohn.hawaii.edu/prestinglab/wiki/INV-Z80-R1-A3 INV-Z80-R1-A3])
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*# BBa_P1010 from pSB1A3 (pSB1A3 from glycerol stock Inventory [http://packrat.stjohn.hawaii.edu/prestinglab/wiki/INV-Z80-R1-A3 INV-Z80-R1-A3])
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*# BBa_P1010 from pSB1A7 (pSB1A7 from glycerol stock Inventory [http://packrat.stjohn.hawaii.edu/prestinglab/wiki/INV-Z80-R1-A3 INV-Z80-R1-A3])
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*# BBa_J33207 from pSB1A2 ([http://partsregistry.org/wiki/index.php/Part:BBa_J33207 BBa_J33207] harboring pSB1A2 from plasmid prep)
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* x2 Controls
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*# positive control: [http://partsregistry.org/wiki/index.php/Part:BBa_K125805 BBa_K125805] from pSB1A3 ([http://partsregistry.org/wiki/index.php/Part:BBa_K125805 BBa_K125805] harboring pSB1A3 from glycerol stock Inventory [http://packrat.stjohn.hawaii.edu/prestinglab/wiki/INV-Z80-R1-D4 INV-Z80-R1-D4])
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*# negative control: H<sub>2</sub>O

Revision as of 09:13, 24 September 2008

Projects Events Resources
Sponsors Experiments Milestones Protocols
Notebook (t) Meetings (t)

Things we did today

Wetlab work

Plasmid prep

Grace
  • pSB1A3
  • rbs+GFPf+tt #1, 2
  • nir+rbs+GFP #1, 4, 7
  • nir+rbs+slr1+GFPf #2, 8
  • nir+rbs+pilA #18

Sequencing

Margaret
1st lane is ladder, 2nd lane is rep amplified from a colony, 3rd lane is rep amplified from a restriction product of rep from the same colony.
  • PCR of rep, ran a gel to verify size, and exo-sap. will send in tomorrow

PCR: pRL1383a MCS Replacement

Norman
  • x6 parallel MCS replacements fragment by PCR amplification. PCR out each MCS replacement insert with HindIII-VF2BB_fx._sb.1 and BamHI-VRBB_rx._sb.1
    1. BBa_I52002 from pSB4A5 (Spring 2008 Plate 1022 1C)
    2. BBa_I52001 from pSB4T5 (Spring 2008 Plate 1020 1A)
    3. BBa_I52002 from BBa_I51020 (BBa_I51020 Base Vector from glycerol stock Inventory [http://packrat.stjohn.hawaii.edu/prestinglab/wiki/INV-Z80-R1-A3 INV-Z80-R1-A3])
    4. BBa_P1010 from pSB1A3 (pSB1A3 from glycerol stock Inventory [http://packrat.stjohn.hawaii.edu/prestinglab/wiki/INV-Z80-R1-A3 INV-Z80-R1-A3])
    5. BBa_P1010 from pSB1A7 (pSB1A7 from glycerol stock Inventory [http://packrat.stjohn.hawaii.edu/prestinglab/wiki/INV-Z80-R1-A3 INV-Z80-R1-A3])
    6. BBa_J33207 from pSB1A2 ([http://partsregistry.org/wiki/index.php/Part:BBa_J33207 BBa_J33207] harboring pSB1A2 from plasmid prep)
  • x2 Controls
    1. positive control: [http://partsregistry.org/wiki/index.php/Part:BBa_K125805 BBa_K125805] from pSB1A3 ([http://partsregistry.org/wiki/index.php/Part:BBa_K125805 BBa_K125805] harboring pSB1A3 from glycerol stock Inventory [http://packrat.stjohn.hawaii.edu/prestinglab/wiki/INV-Z80-R1-D4 INV-Z80-R1-D4])
    2. negative control: H2O


Discussion

Quote of the Day

History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson


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