Paris/July 25

From 2007.igem.org

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|Backward insert - EcoRI site still presents : we can't use EcoRI
|Backward insert - EcoRI site still presents : we can't use EcoRI
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== Ligation & transformation reactions ==
 +
 +
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The Ligation reactions performed yesterday have all failed (the reactions were performed at 42° by mistake, then, another 1µL of ligase was added for a 5min rxn at 25°)
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The following reactions were performed today
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 +
{|
 +
|- style="background: #ccccff;"
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! colspan="5" style="background: #ccccff;" | Ligations
 +
|- style="background: #ccccff; text-align: center;"
 +
|width=5%| Number
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|width=25%| Insert
 +
|width=5%| Insert Volume (µL)
 +
|width=25%| Vector
 +
|width=5%| Vector Volume (µL)
 +
|width=35%| Comments
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |L1
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|D23.1 (AraC/pBad promoter FI)
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|8.5
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|D9.1 (J61002 ready for insertion of a FI)
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|1.5
 +
|
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |L2
 +
|D18 (BB dig. lox66DapAE.coli)
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|7
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|D22.1 (pSB1A2 Eco, Pst)
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|3
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|
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|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |L3
 +
|D19 (BB dig. lox66DapAsubtilis)
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|7
 +
|D22.1 (pSB1A2 Eco, Pst)
 +
|3
 +
|
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|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |L4
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|D20 (Lox66-DapAE.coli BI)
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|8
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|D1 (pJ23100 BV)
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|2
 +
|
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|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |L5
 +
|D21 (lox66-DapAsubtilis BI)
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|6.5
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|D1 (pJ23100 BV)
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|2 (+1.5µL H2O)
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|
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |L6
 +
|D14.1 (Cre ORF)
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|8
 +
|D15.1 (B0030 BV)
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|2
 +
|
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |L7
 +
|D27 (Lox71-ftsZ BI)
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|6
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|D1 (pJ23100 BV)
 +
|2 (+2µL H2O)
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|
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|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |L8
 +
|D27 (Lox71-ftsZ BI)
 +
|6
 +
|D3 (pJ23107 BV)
 +
|2 (+2 H2O)
|
|
|}
|}

Revision as of 16:05, 25 July 2007

Digestions

Digestion of purifications of PCR products

Digestion Products
Number Product Name Matrix Name Enzyme 1 Enzyme 2 size Description
D28 RBS-DapAColi BB P8 (RBS-DapAColi) EcoRI PstI ~950 Biobrick
D29 RBS-DapAColi P8 (RBS-DapAColi) XbaI PstI ~950 Backward insert
D30 RBS-DapASubtilis BB P9 (RBS-DapASubtilis) EcoRI PstI ~950 Biobrick
D31 RBS-DapASubtilis P9 (RBS-DapASubtilis) XbaI PstI ~950 Backward insert
D32 Lox71-FtsA-FtsZ P10 Lox71-FtsA-FtsZ XbaI PstI ~2600 Backward insert - EcoRI site still presents : we can't use EcoRI


Ligation & transformation reactions

The Ligation reactions performed yesterday have all failed (the reactions were performed at 42° by mistake, then, another 1µL of ligase was added for a 5min rxn at 25°)

The following reactions were performed today

Ligations
Number Insert Insert Volume (µL) Vector Vector Volume (µL) Comments
L1 D23.1 (AraC/pBad promoter FI) 8.5 D9.1 (J61002 ready for insertion of a FI) 1.5
L2 D18 (BB dig. lox66DapAE.coli) 7 D22.1 (pSB1A2 Eco, Pst) 3
L3 D19 (BB dig. lox66DapAsubtilis) 7 D22.1 (pSB1A2 Eco, Pst) 3
L4 D20 (Lox66-DapAE.coli BI) 8 D1 (pJ23100 BV) 2
L5 D21 (lox66-DapAsubtilis BI) 6.5 D1 (pJ23100 BV) 2 (+1.5µL H2O)
L6 D14.1 (Cre ORF) 8 D15.1 (B0030 BV) 2
L7 D27 (Lox71-ftsZ BI) 6 D1 (pJ23100 BV) 2 (+2µL H2O)
L8 D27 (Lox71-ftsZ BI) 6 D3 (pJ23107 BV) 2 (+2 H2O)
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