From 2007.igem.org

Contents 1 PCRs 2 Glycerol stock 3 Minipreps on Biobricks 4 E.Coli pKS::DGAT

PCRs

PCR : Lox71-KmR-Lox66
PCR Settings		Buffer (5x)	5x 10µL			Expected size
Annealing (°C)		MgCl2 10µM	10µM 0µL			1084
55		dNTP 10µM	10µM 1µL			Success
Time Elongation		Oligo F 10µM	10 Lox71-KmR-F	2.5µL		?
3m00'		Oligo R 10µM	11 Lox66-KmR-R	2.5µL		Image (click to enlarge)
Number cycles		Water	34µl			[[Image:|30px]]
35		Polymerase	Phusion 0.5µL			Band (0=ladder)
		DNA	MP3_pSB1AK3-BBa_B0015 0.5µl

PCR : Lox71-FtsA-FtsZ
PCR Settings		Buffer (5x)	5x 10µL			Expected size
Annealing (°C)		MgCl2 10µM	10µM 0µL			2588
55		dNTP 10µM	10µM 1µL			Success
Time Elongation		Oligo F 10µM	3 Lox71-FtsA-F	2.5µL		? The PCR seemed to have worked but the purification failed
3m00'		Oligo R 10µM	2 FtsZ-R	2.5µL		Image (click to enlarge)
Number cycles		Water	34µl			[[Image:|30px]]
35		Polymerase	Phusion 0.5µL			Band (0=ladder)
		DNA	toothpick in MG1655 glycerol

Glycerol stock

• BBa_pJ23107 clone 3&4 (S14)
• BBa_I0500 clone 1&2 (S13)

Minipreps on Biobricks

• BBa_pJ23107 clone 3&4
• BBa_I0500 clone 1&2

E.Coli pKS::DGAT

We look under microscopy 36 hours after incubation of E.coli transformed by pKS::DGAT and the control E.coli transformed by part B0015 on different LB medium (See July 18).

• Observations:

We can observe colonies (X10).

• Interpretation

We can observe lipid inclusions within colonies with DGAT. Further investigations need single cell visualisation.