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Revision as of 13:02, 30 July 2007 (view source) Eismoustique (Talk | contribs) (→DIgestion reactions) ← Older edit		Revision as of 14:24, 30 July 2007 (view source) Eismoustique (Talk | contribs) (→DIgestion reactions) Newer edit →
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

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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