Paris/July 25

From 2007.igem.org

(Difference between revisions)
(Ligation & transformation reactions)
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|D3 (pJ23107 BV)
|D3 (pJ23107 BV)
|2 (+2 H2O)
|2 (+2 H2O)
 +
|
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |L9
 +
|lox66 [O14+O15] (0.2µM)
 +
|2
 +
|D22.1 (pSB1A2 Eco, Pst)
 +
|2 (+6 H2O)
 +
|
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |L10
 +
|lox71 [O16+O17] (0.2µM)
 +
|2
 +
|D22.1 (pSB1A2 Eco, Pst)
 +
|2 (+6 H2O)
|
|
|}
|}

Revision as of 16:20, 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)
L9 lox66 [O14+O15] (0.2µM) 2 D22.1 (pSB1A2 Eco, Pst) 2 (+6 H2O)
L10 lox71 [O16+O17] (0.2µM) 2 D22.1 (pSB1A2 Eco, Pst) 2 (+6 H2O)