Paris/August 20

From 2007.igem.org

< Paris(Difference between revisions)
(Colony PCR)
 
(One intermediate revision not shown)
Line 123: Line 123:
|D34 (lox66 BV)
|D34 (lox66 BV)
|4
|4
-
|lox66-RBS-dapAcoli BI
+
|lox66-RBS-dapAcoli  
| +++
| +++
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
Line 131: Line 131:
|D34 (lox66 BV)
|D34 (lox66 BV)
|4
|4
-
|lox66-RBS-dapAsubtilis BI
+
|lox66-RBS-dapAsubtilis  
|RAS
|RAS
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
Line 139: Line 139:
|D22 (pSB1A2 open vector EP)
|D22 (pSB1A2 open vector EP)
|4µl
|4µl
-
|RBS-dapAcoli cloned in pSB1A2
+
|RBS-dapAcoli cloned in <bbpart>pSB1A2</bbpart>
| +++
| +++
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
Line 147: Line 147:
|D22 (pSB1A2 open vector EP)
|D22 (pSB1A2 open vector EP)
|4µl
|4µl
-
|RBS-dapAsubtilis cloned in pSB1A2
+
|RBS-dapAsubtilis cloned in <bbpart>pSB1A2</bbpart>
| +
| +
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
Line 155: Line 155:
|D47 (<bbpart>pSB1A2</bbpart> XP dig.)
|D47 (<bbpart>pSB1A2</bbpart> XP dig.)
|4µl
|4µl
-
|lox71-ftsZ cloned in pSB1A2
+
|lox71-ftsZ cloned in <bbpart>pSB1A2</bbpart>
| +
| +
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
Line 163: Line 163:
|D47 (<bbpart>pSB1A2</bbpart> XP dig.)
|D47 (<bbpart>pSB1A2</bbpart> XP dig.)
|4µl
|4µl
-
|lox71-ftsA-Z cloned in pSB1A2
+
|lox71-ftsA-Z cloned in <bbpart>pSB1A2</bbpart>
| +
| +
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
Line 171: Line 171:
|D48 (<bbpart>pSB1A2</bbpart> ES dig.)
|D48 (<bbpart>pSB1A2</bbpart> ES dig.)
|2µl
|2µl
-
|RBS-dapAsubtilis cloned in pSB1A2
+
|RBS-dapAsubtilis cloned in <bbpart>pSB1A2</bbpart>
| +
| +
|}
|}
Line 187: Line 187:
*L42
*L42
*L43
*L43
 +
 +
 +
== Sequencing ==
 +
 +
The following sequencing reactions have been performed
 +
 +
TL_MPT5.1+O41
 +
TL_MPT5.2+O41
 +
TL_L26.1+O18
 +
TL_L26.1+O19
 +
TL_L26.2+O18
 +
TL_L26.2+O19
 +
TL_L27.2+O18
 +
TL_L27.2+O19
 +
TL_L27.3+O18
 +
TL_L27.3+O19
 +
TL_L27.4+O18
 +
TL_L27.4+O19
 +
TL_L27.5+O18
 +
TL_L27.5+O19
 +
TL_L28.2+O18
 +
TL_L28.2+O19
 +
TL_L28.3+O18
 +
TL_L28.3+O19
 +
TL_L28.8+O18
 +
TL_L28.8+O19
 +
TL_L29.1+O18
 +
TL_L29.1+O19
 +
TL_L29.2+O18
 +
TL_L29.2+O19
 +
TL_L30A1+O18
 +
TL_L30A1+O19
 +
TL_L30B1+O18
 +
TL_L30B1+O19
 +
TL_L27.2+O24
 +
TL_L27.3+O24
 +
TL_L27.4+O24
 +
TL_L27.5+O24
 +
TL_L28.2+O24
 +
TL_L28.3+O24
 +
TL_L28.8+O24
 +
TL_L28.2+O22
 +
TL_L28.3+O22
 +
TL_L28.8+O22
 +
TL_L28.2+O23
 +
TL_L28.3+O23
 +
TL_L28.8+O23

Latest revision as of 14:48, 30 September 2007

yesterday -- tomorrow

Ligations

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 +++
L38 D54 (RBS-dapAsubtilis BI) 13 D34 (lox66 BV) 4 lox66-RBS-dapAsubtilis 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 +

5 points with :

  • L31
  • L32
  • L34
  • L35
  • L36
  • L37
  • L39
  • L40 (2 points only)
  • L41 (2 points only)
  • L42
  • L43


Sequencing

The following sequencing reactions have been performed

TL_MPT5.1+O41 TL_MPT5.2+O41 TL_L26.1+O18 TL_L26.1+O19 TL_L26.2+O18 TL_L26.2+O19 TL_L27.2+O18 TL_L27.2+O19 TL_L27.3+O18 TL_L27.3+O19 TL_L27.4+O18 TL_L27.4+O19 TL_L27.5+O18 TL_L27.5+O19 TL_L28.2+O18 TL_L28.2+O19 TL_L28.3+O18 TL_L28.3+O19 TL_L28.8+O18 TL_L28.8+O19 TL_L29.1+O18 TL_L29.1+O19 TL_L29.2+O18 TL_L29.2+O19 TL_L30A1+O18 TL_L30A1+O19 TL_L30B1+O18 TL_L30B1+O19 TL_L27.2+O24 TL_L27.3+O24 TL_L27.4+O24 TL_L27.5+O24 TL_L28.2+O24 TL_L28.3+O24 TL_L28.8+O24 TL_L28.2+O22 TL_L28.3+O22 TL_L28.8+O22 TL_L28.2+O23 TL_L28.3+O23 TL_L28.8+O23