Paris/August 20
From 2007.igem.org
(Difference between revisions)
(→Colony PCR) |
|||
| Line 74: | Line 74: | ||
|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 82: | ||
|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 98: | Line 98: | ||
|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 106: | ||
|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 114: | ||
|4 | |4 | ||
|lox66-mRFP | |lox66-mRFP | ||
| - | |+++ | + | | +++ |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
| style="background: #ccffcc;" |L37 | | style="background: #ccffcc;" |L37 | ||
| Line 122: | Line 122: | ||
|4 | |4 | ||
|lox66-RBS-dapAcoli BI | |lox66-RBS-dapAcoli BI | ||
| - | |+++ | + | | +++ |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
| style="background: #ccffcc;" |L38 | | style="background: #ccffcc;" |L38 | ||
| Line 138: | Line 138: | ||
|4µl | |4µl | ||
|RBS-dapAcoli cloned in pSB1A2 | |RBS-dapAcoli cloned in pSB1A2 | ||
| - | |+++ | + | | +++ |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
| style="background: #ccffcc;" |L40 | | style="background: #ccffcc;" |L40 | ||
| Line 146: | Line 146: | ||
|4µl | |4µl | ||
|RBS-dapAsubtilis cloned in pSB1A2 | |RBS-dapAsubtilis cloned in pSB1A2 | ||
| - | |+ | + | | + |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
| style="background: #ccffcc;" |L41 | | style="background: #ccffcc;" |L41 | ||
| Line 154: | Line 154: | ||
|4µl | |4µl | ||
|lox71-ftsZ cloned in pSB1A2 | |lox71-ftsZ cloned in pSB1A2 | ||
| - | |+ | + | | + |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
| style="background: #ccffcc;" |L42 | | style="background: #ccffcc;" |L42 | ||
| Line 162: | Line 162: | ||
|4µl | |4µl | ||
|lox71-ftsA-Z cloned in pSB1A2 | |lox71-ftsA-Z cloned in pSB1A2 | ||
| - | |+ | + | | + |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
| style="background: #ccffcc;" |L43 | | style="background: #ccffcc;" |L43 | ||
| Line 170: | Line 170: | ||
|2µl | |2µl | ||
|RBS-dapAsubtilis cloned in pSB1A2 | |RBS-dapAsubtilis cloned in pSB1A2 | ||
| - | |+ | + | | + |
|} | |} | ||
Revision as of 12:58, 20 August 2007
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 | + |
