Paris/August 20

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Colony PCR

Ligations
Number Insert Insert Volume (µL) Vector Vector Volume (µL) Comments Number of colonies
C1 MP3.1 digested with EcoRI (D0) 2 control of ligation efficiency
C3 D34 (lox66 BV) 4µl control of D34 digestion
C5 D50 (BBa_J61002 BV) 1.5µl control of D50 digestion
C6 D41 (araC/pBad-J61002 plasmid SP dig.) 1µl control of D41 digestion
C7 D22 (pSB1A2 open vector EP) 4µl control of D22 digestion
C8 D47 (pSB1A2 XP dig.) 4µl control of D47 digestion
C9 D48 (pSB1A2 ES dig.) 2µl control of D48 digestion
L31 D44 (B0015 terminator BI) 4 D35 (lox71 BV) 3 Insertion of B0015 terminator behind lox71
++
L32 D45 (B0015 terminator FI) 4 D42 (Lox66 FV) 3 Insertion of B0015 terminator in front of lox66
++
L33 D33.1 (RBS(B0030)CreORF BI) 5 D50 (BBa_J61002 BV) 1.5 pTet>>RBS-Cre RAS
L34 D49 (lox71-gfptripart BI) 5 D50 (BBa_J61002 BV) 1.5 pTet>>lox71-gfptripart
+
L35 D49 (lox71-gfptripart BI) 5 D41 (araC/pBad-J61002 plasmid SP dig.) 1 araC/pBad>>lox71-gfptripart
++
L36 D13.4 (mRFP dig. SP) 6 D34 (lox66 BV) 4 lox66-mRFP
++
L37 D52 (RBS-dapAcoli BI) 6 D34 (lox66 BV) 4 lox66-RBS-dapAcoli BI
++
L38 D54 (RBS-dapAsubtilis BI) 13 D34 (lox66 BV) 4 lox66-RBS-dapAsubtilis BI RAS
L39 D51 (RBS-dapAcoli BBinsert) 4 D22 (pSB1A2 open vector EP) 4µl RBS-dapAcoli cloned in pSB1A2
++
L40 D53 (RBS-dapAsubtilis BBinsert) 13 ( D22 (pSB1A2 open vector EP) 4µl RBS-dapAsubtilis cloned in pSB1A2
L41 D27' (lox71-ftsZ bw-insert) 12 D47 (pSB1A2 XP dig.) 4µl lox71-ftsZ cloned in pSB1A2
L42 D32 (lox71-ftsAftsZ bw-insert) 6 D47 (pSB1A2 XP dig.) 4µl lox71-ftsA-Z cloned in pSB1A2
L43 D55 (RBS-dapAsubtilis FI) 15 D48 (pSB1A2 ES dig.) 2µl RBS-dapAsubtilis cloned in pSB1A2