Paris/August 20
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- | == | + | [[Paris/August 19|yesterday]] -- [[Paris/August 21|tomorrow]] |
+ | |||
+ | == Ligations == | ||
{| | {| | ||
|- style="background: #ccccff;" | |- style="background: #ccccff;" | ||
Line 74: | Line 76: | ||
|3 | |3 | ||
| Insertion of B0015 terminator behind lox71 | | Insertion of B0015 terminator behind lox71 | ||
- | | | + | | +++ |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
| style="background: #ccffcc;" |L32 | | style="background: #ccffcc;" |L32 | ||
Line 82: | Line 84: | ||
|3 | |3 | ||
| Insertion of B0015 terminator in front of lox66 | | Insertion of B0015 terminator in front of lox66 | ||
- | | | + | | +++ |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
| style="background: #ccffcc;" |L33 | | style="background: #ccffcc;" |L33 | ||
Line 90: | Line 92: | ||
|1.5 | |1.5 | ||
| pTet>>RBS-Cre | | pTet>>RBS-Cre | ||
- | | | + | |RAS |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
| style="background: #ccffcc;" |L34 | | style="background: #ccffcc;" |L34 | ||
Line 98: | Line 100: | ||
|1.5 | |1.5 | ||
|pTet>>lox71-gfptripart | |pTet>>lox71-gfptripart | ||
- | | | + | | ++ |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
| style="background: #ccffcc;" |L35 | | style="background: #ccffcc;" |L35 | ||
Line 106: | Line 108: | ||
|1 | |1 | ||
|araC/pBad>>lox71-gfptripart | |araC/pBad>>lox71-gfptripart | ||
- | | | + | | +++ |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
| style="background: #ccffcc;" |L36 | | style="background: #ccffcc;" |L36 | ||
Line 114: | Line 116: | ||
|4 | |4 | ||
|lox66-mRFP | |lox66-mRFP | ||
- | | | + | | +++ |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
| style="background: #ccffcc;" |L37 | | style="background: #ccffcc;" |L37 | ||
Line 121: | Line 123: | ||
|D34 (lox66 BV) | |D34 (lox66 BV) | ||
|4 | |4 | ||
- | |lox66-RBS-dapAcoli | + | |lox66-RBS-dapAcoli |
- | | | + | | +++ |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
| style="background: #ccffcc;" |L38 | | style="background: #ccffcc;" |L38 | ||
Line 129: | Line 131: | ||
|D34 (lox66 BV) | |D34 (lox66 BV) | ||
|4 | |4 | ||
- | |lox66-RBS-dapAsubtilis | + | |lox66-RBS-dapAsubtilis |
- | | | + | |RAS |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
| style="background: #ccffcc;" |L39 | | style="background: #ccffcc;" |L39 | ||
Line 137: | 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;" | ||
| style="background: #ccffcc;" |L40 | | style="background: #ccffcc;" |L40 | ||
Line 145: | 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;" | ||
| style="background: #ccffcc;" |L41 | | style="background: #ccffcc;" |L41 | ||
Line 153: | 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;" | ||
| style="background: #ccffcc;" |L42 | | style="background: #ccffcc;" |L42 | ||
Line 161: | 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;" | ||
| style="background: #ccffcc;" |L43 | | style="background: #ccffcc;" |L43 | ||
Line 169: | 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> |
- | | | + | | + |
|} | |} | ||
+ | |||
+ | 5 points with : <br> | ||
+ | *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 |
Latest revision as of 14:48, 30 September 2007
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