Paris/July 30

From 2007.igem.org

(Difference between revisions)
(DIgestion reactions)
(DIgestion reactions)
Line 66: Line 66:
| style="background: #ccffcc;" |D33
| style="background: #ccffcc;" |D33
|RBS(B0030)CreORF
|RBS(B0030)CreORF
-
|L6 (clone 1 & 2)
+
|L6' (clone 1 & 2)
|XbaI
|XbaI
|PstI
|PstI
Line 76: Line 76:
| style="background: #ccffcc;" |D34
| style="background: #ccffcc;" |D34
|lox66
|lox66
-
|L9 lox66
+
|L9' lox66
|SpeI
|SpeI
|PstI
|PstI
Line 86: Line 86:
| style="background: #ccffcc;" |D35
| style="background: #ccffcc;" |D35
|lox71
|lox71
-
|L10 lox71
+
|L10' lox71
|SpeI
|SpeI
|PstI
|PstI

Revision as of 14:24, 30 July 2007

Colonies PCR screen

On colony PCRs performed to screen for correct ligation reactions (performed on 27/07/07):

10 clones of L1 ligation reactions, 6 clones of L6 ligation reactions

preparation of the following PCR mix

  • 18x (12.5µL 2xreaction mix)= 225
  • 18x (0.5µL oligo O18 VF2 10µM)=9
  • 18x (0.5µL oligo O19 VR 10µM)=9
  • 12µL H20+colony (10min 100°)

PCRs

DIgestion reactions

The following digestion reactions have been performed:

Digestion Products
Number Product Name Matrix Name Enzyme 1 Enzyme 2 size Description
D19' BB dig. lox66DapAsubtilis lox66-DapAsubtilis P6' PCR product EcoRI PstI ~960 PCR product digested for cloning as a BioBrick
D21' bw-insert lox66-DapAsubtilis lox66-DapAsubtilis P6' PCR product XbaI PstI ~960 PCR product digested for cloning as a BI
D27' lox71-ftsZ bw-insert lox71-ftsZ PCR product P7 XbaI PstI lox71-ftsZ(unmutated) ready for use as a backward insert
D33 RBS(B0030)CreORF L6' (clone 1 & 2) XbaI PstI RBS-Cre digestion for use as a BI
D34 lox66 L9' lox66 SpeI PstI Biobrick plasmid carrying lox66 digested as a BV
D35 lox71 L10' lox71 SpeI PstI Biobrick plasmid carrying lox71 digested as a BV