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Line 63:		Line 63:
	|		|
	|- style="background: #cccccc;"		|- style="background: #cccccc;"
-	| style="background: #ccffcc;" |D3...	+	| style="background: #ccffcc;" |D33
	|RBS(B0030)CreORF		|RBS(B0030)CreORF
	|L6 (clone 1 & 2)		|L6 (clone 1 & 2)
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	|		|
	|- style="background: #cccccc;"		|- style="background: #cccccc;"
-	| style="background: #ccffcc;" |D3...	+	| style="background: #ccffcc;" |D34
	|lox66		|lox66
	|L9 lox66		|L9 lox66
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	|		|
	|- style="background: #cccccc;"		|- style="background: #cccccc;"
-	| style="background: #ccffcc;" |D3...	+	| style="background: #ccffcc;" |D35
	|lox71		|lox71
	|L10 lox71		|L10 lox71

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Revision as of 12:59, 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
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