Team:Aalto-Helsinki/LabBook

• Project
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  • Models of propane pathway
    • Kinetics of propane pathway
    • Model of CAR activation
  • Synergy model
  • Micelle model
  • About cellulose pathway
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• Week 1

• Week 2

• Week 3

• Week 4

• Week 5

• Week 6

• Week 7

• Week 8

• Week 9

• Week 10

• Week 11

• Week 12

• Week 13

• Week 14

• Week 15

• Week 16

• Week 17

• Week 18

Lab Book

An example of what the table could look like:

Construct worked with	Things done						Lab 1.1.2015 Monday
CAR	Here is my text	Here is my text	Here is my text	Here is my text	Here is my text	Here is my text	Here is my text, there is lot of it
another CAR?	Here is my text	Here is my text	Here is my text	Here is my text	Here is my text	Here is my text	Here is my text, there is lot of it
Ado?	Here is my text	Here is my text	Here is my text	Here is my text	Here is my text
Ado cont.			Here is my text	Here is my text	Here is my text	Here is my text	Here is my text, there is lot of it

Week 1

Construct worked with	Things done						May 18th - May 22nd
Preparations Plates	Making Cm plates	Stored in +4°C
Preparations Competent Cells	Preparing solutions for competent cells	Stored in +4°C
	o/n culture of E. coli TOP10	Chemical competent cells protocol	Tranformation efficiency kit
	o/n culture of E. coli BL21(DE3)	Chemical competent cells protocol	Tranformation efficiency kit	Colonies are the wrong color
Preparations Propagations	Transformed   AH001-AH008 into TOP10	AH001-AH004 & AH006	Stored in +4°C
		No colonies for    AH005, AH007 or AH008

Week 2

Construct worked with	Things done						May 25th - May 29th
Preparations Propagations	Transformed AH007-AH009, AH005 & AH019 into TOP10	AH007-AH009 & AH019	o/n cultures	MiniPrep
	o/n culture of  AH001-AH004 & AH006	MiniPrep
		No colonies for AH005	Re-transform into TOP10	Plates in rt over weekend
Checking Competent Cells	o/n culture of BL21(DE3) transformants	MiniPrep	PCR with VF2 & VR	Smeared gels
			AH001+    AH003+AH008 -> AH013	Repeat ligation	Restrict & extract from gel	Transformation into TOP10	Plates in rt over weekend
Testing RBS's Assembly	Restrict AH001-AH004 & AH006-AH008	Ligation	AH002+   AH004+AH008 -> AH014	Repeat ligation	Restrict & extract from gel	Transformation into TOP10	Plates in rt over weekend
			AH001+    AH006+AH008 -> AH011	Repeat ligation	Restrict & extract from gel	Transformation into TOP10	Plates in rt over weekend

Week 3

Construct worked with	Things done						June 1st - June 5th
Preparations Propagations	Transform     AH020 & AH021 into TOP10	Plates in rt  over weekend
Testing RBS's Assembly	No colonies for AH011 or AH014	re-assemble	Restrict & extract from gel	Ligate & tranform into TOP10	No colonies
	AH005 & AH013 OK	o/n cultures	MiniPrep

Week 4

Construct worked with	Things done						June 8th - June 12th
Preparations Propagations	o/n cultures for AH020 & AH021	MiniPrep
Testing RBS's Assembly	Re-assembling   AH011, AH013 & AH014	Transform into TOP10	o/n cultures	MinPrep
	Assembling AH015	Ligate AH002+   AH005+AH008	Restrict & extract from gel	Transform into TOP10	o/n cultures	MiniPrep

Week 5

Week 6

Construct worked with	Things done						June 22nd - June 26th
Testing RBS's Assembly	Re-assembling AH016 & AH018	Transform into BL21(DE3)	No colonies
Testing RBS's Propagation	o/n cultures of  AH016 & AH018 in TOP10	MiniPrep	Transform into BL21(DE3)	Plates in rt over the weekend
Testing RBS's Measurement	AH018 in BL21(DE3)	o/n culture	IPTG induction &     blue chromoprotein measurement	No blue chromoprotein
		Refresh on IPTG plate	No blue chromoprotein
Preparations Linearized Backbones	o/n cultures of AH007-AH009	MiniPrep	Restrict & extract from gel

Week 7

Construct worked with	Things done						June 29th - July 3rd
Preparations Linearized Backbones	Transform AH007-AH009 into TOP10	No colonies (mistake in protocol)	Use iGEM's linearized backbones
Testing RBS's	o/n cultures of   AH016&AH018 in TOP10	IPTG induction	No Blue chromoprotein
Testing RBS's Measurement	Refresh AH016 & AH018 on IPTG plate	No blue chromoprotein
Violacein Plasmid Construction	PCR AH20 with P012&P013	PCR purification	VioA waits for 2nd PCR
	PCR AH20 with P024&P025	PCR unsuccesful
	PCR AH20 with P034&P035	PCR purification	VioE waits for 2nd PCR

Week 8

Construct worked with	Things done						July 6th - July 10th
Preparations	Preparing   PK BL21(DE3) competent cells	Cells stored in -80°C
Cellulose Plasmid Construction	PCR AH021 with P019&P020	Problem with PCR machine	Re-do PCR	PCR purifcation	BglX waits for 2nd PCR
	AH029 into TOP10	Transformants stored in +4°C
Testing RBS's	Transform AH016 & AH018 into PK BL21(DE3)	Refresh transformants on IPTG plates	No blue chromoprotein
Violacein Plasmid Construction	Gradient PCR for AH020 with P024&P025	PCR purification

Week 9

Construct worked with	Things done						July 13th - July 17th
Preparations Propagations	o/n culture of AH029	MiniPrep
		CAR2 OK	PCR purification
Propane Plasmids 1&2 Construction	PCR for CAR1-3 & AtoB1-2	CAR1,3 & AtoB1-2 unspecific bands	Gradient PCR for CAR3 & 4
		PCR with DMSO for CAR1, 3 & AtoB1, 2	dunnowhat
GFP Brick Construction	o/n culture of AH044	MiniPrep
Propane Plasmid 2 Construction	PCR for AtoB3-4	Unspecific bands	AtoB 4 extracted from gel	Low concentration
		Gradient PCR for AtoB3	Extract & purify from gel

Week 10

Week 11

Construct worked with	Things done						July 27th - July 31st
Propane Plasmid 1 Construction	Continue CAR1 colony screening	Screen more colonies	PCR purification
	PCR for new CAR2	Gradient PCR with KAPA polymerase	PCR purification
Preparations Linearized Backbones	Re-restrict AH043	Gel purification	Low yield	Re-restrict
Propagations	o/n culture of AH031	MiniPrep
	Transform AH033 into TOP10	o/n culture	MiniPrep	Low yield	o/n culture	MiniPrep	Low yield, but stored in -20°C
Propane Plasmid 2 Construction	TOPO Blunt End Cloning for AtoB1 and transformation into TOP10	Screening with colony PCR	PCR purification
Cellulose Plasmid Construction	PCR for Cen1 & Cen2	Gradient PCR with KAPA polymerase	PCR purification for Cen2
			Gel purification for Cen1
	2nd PCR for BglX (from AH021)	Stored in -4°C over the weekend
Propane Plasmid 1 Construction	Re-do colony PCR for CAR3 successful TOPO colony	Re-do colony PCR
		o/n culture of the colony	MiniPrep	PCR
Violacein Plasmid Construction	2nd PCR for VioA (from AH020)	Stored in -4°C over the weekend
	KAPA gradient PCR for AtoB3,4 & CAR3	Gel purification for AtoB4 & Car3
		PCR purification for AtoB3

Week 12

Week 13

Week 14

Week 15

Week 16

Week 17

Week 18