Paris/July 30

From 2007.igem.org

(Difference between revisions)
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== PCRs ==
== PCRs ==
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== DIgestion reactions ==
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The following digestion reactions have been performed:
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{|
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|- style="background: #ccccff;"
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! colspan="5" style="background: #ccccff;" | Digestion Products
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|- style="background: #ccccff; text-align: center;"
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|width=5%| Number
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|width=20%| Product Name
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|width=30%| Matrix Name
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|width=5%| Enzyme 1
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|width=5%| Enzyme 2
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|width=5%| size
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|width=35%| Description
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|- style="background: #cccccc;" 
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| style="background: #ccffcc;" |D19'
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|BB dig. lox66DapAsubtilis
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|lox66-DapAsubtilis P6' PCR product
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|EcoRI
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|PstI
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|~960
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|PCR product digested for cloning as a BioBrick
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|
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|
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|- style="background: #cccccc;" 
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| style="background: #ccffcc;" |D21'
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|bw-insert lox66-DapAsubtilis
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|lox66-DapAsubtilis P6' PCR product
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|XbaI
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|PstI
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|~960
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|
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|
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|- style="background: #cccccc;" 
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| style="background: #ccffcc;" |D27'
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|lox71-ftsZ bw-insert
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|lox71-ftsZ PCR product P7
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|XbaI
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|PstI
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|
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|lox71-ftsZ(unmutated) ready for use as a backward insert
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|
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|
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|- style="background: #cccccc;" 
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| style="background: #ccffcc;" |D3...
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|RBS(B0030)CreORF
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|L6 (clone 1 & 2)
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|XbaI
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|PstI
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|
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|RBS-Cre digestion for use as a BI
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|
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|
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|- style="background: #cccccc;" 
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| style="background: #ccffcc;" |D3...
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|lox66
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|L9 lox66
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|SpeI
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|PstI
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|
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|Biobrick plasmid carrying lox66 digested as a BV
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|
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|
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|- style="background: #cccccc;" 
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| style="background: #ccffcc;" |D3...
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|lox71
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|L10 lox71
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|SpeI
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|PstI
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|
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|Biobrick plasmid carrying lox71 digested as a BV
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|
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|
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|}

Revision as of 12:55, 30 July 2007

Colonies PCR screen

On colony PCRs performed to screen for correct ligation reactions (performed on 27/07/07):

10 clones of L1 ligation reactions, 6 clones of L6 ligation reactions

preparation of the following PCR mix

  • 18x (12.5µL 2xreaction mix)= 225
  • 18x (0.5µL oligo O18 VF2 10µM)=9
  • 18x (0.5µL oligo O19 VR 10µM)=9
  • 12µL H20+colony (10min 100°)

PCRs

DIgestion reactions

The following digestion reactions have been performed:

Digestion Products
Number Product Name Matrix Name Enzyme 1 Enzyme 2 size Description
D19' BB dig. lox66DapAsubtilis lox66-DapAsubtilis P6' PCR product EcoRI PstI ~960 PCR product digested for cloning as a BioBrick
D21' bw-insert lox66-DapAsubtilis lox66-DapAsubtilis P6' PCR product XbaI PstI ~960
D27' lox71-ftsZ bw-insert lox71-ftsZ PCR product P7 XbaI PstI lox71-ftsZ(unmutated) ready for use as a backward insert
D3... RBS(B0030)CreORF L6 (clone 1 & 2) XbaI PstI RBS-Cre digestion for use as a BI
D3... lox66 L9 lox66 SpeI PstI Biobrick plasmid carrying lox66 digested as a BV
D3... lox71 L10 lox71 SpeI PstI Biobrick plasmid carrying lox71 digested as a BV
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