Revision as of 09:25, 28 August 2015

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
To the Top
To the Previous Page

Lab Book

Week 1

Worked With	WorkFlow				May 18th - May 22nd
Preparations Plates	Making Cm plates	Stored in +4°C
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
Propagations	Transformed AH001-AH008 into TOP10	AH001-AH004 & AH006	Stored in +4°C
		No colonies for AH005, AH007 or AH008

Week 2

Worked With	WorkFlow						May 25th - May 29th
Preparations Propagating	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	Transform 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	Transform into TOP10	Plates in rt over weekend
			AH001+ AH006+AH008 -> AH011	Repeat ligation	Restrict & extract from gel	Transform into TOP10	Plates in rt over weekend

Week 3

Worked With	WorkFlow						June 1st - June 5th
Preparations Propagating	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
	Assembling AH015	Ligate AH002+ AH005+AH008	Restrict & extract from gel	Transform into TOP10	o/n cultures	No colonies on the plate

Week 4

Worked With	WorkFlow						June 8th - June 12th
Preparations Propagating	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

Worked With	WorkFlow						June 15th - June 19th
						AH016 wrong size on gel
				Colonies in BL21(DE3)	o/n cultures & MiniPrep	AH018 OK on gel
Testing RBS's Assembly	Screening AH011 & AH013-AH015 by gel electrophoresis	AH011, AH013 & AH015 OK	Assembling AH016 & AH018	Transform into TOP10 & BL21(DE3)	T7 promoter doesn't work in TOP10...
		AH014 incorrect size	Assembling AH017 with multiple AH014 plasmids	Restrict & extract from gel	Transform into BL21(DE3)	Colonies are not blue
	Re-assembling AH016 & AH018	Restrict extract from gel	Transform into BL21(DE3)	Colonies are not blue

Preparations Linearized Backbones	o/n cultures of AH007-AH009	MiniPrep	Restrict & extract from gel	Low concentration

Week 6

Worked With	WorkFlow				June 22nd - June 26th
Preparations Linearized Backbones	o/n cultures of AH007-AH009	MiniPrep	Restrict & extract from gel

Testing RBS's Assembly	Re-assembling AH016 & AH018	Transform into BL21(DE3)	No colonies
Propagation	o/n cultures of AH016 & AH018 in TOP10	MiniPrep	Transform into BL21(DE3)	Plates in rt over the weekend
Measurement	AH018 in BL21(DE3)	o/n culture	IPTG induction & blue chromoprotein measurement	No blue chromoprotein
		Refresh on IPTG plate	No blue chromoprotein

Week 7

Worked With	WorkFlow			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 BL21	IPTG induction	No Blue chromoprotein
	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

Worked With	WorkFlow					July 6th - July 10th
Preparations Competent Cells	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

Worked With	WorkFlow					July 13th - July 17th
Preparations Propagations	o/n culture of AH029	MiniPrep

	CAR2 OK	PCR purification	An illegal restriction site found in the sequence
Propane Plasmid 1 Construction	PCR for CAR1-3	CAR1 & 3 unspecific bands	Gradient PCR for CAR3	PCR with DMSO for CAR3	Store in -20°C
		PCR with DMSO for CAR1	No product

	PCR with DMSO for AtoB1	Store in -20°C
Propane Plasmid 2 Construction	PCR for AtoB 1-4	Gradient PCR for AtoB2	PCR with DMSO for AtoB2	Store in -20°C
	Gradient PCR for AtoB3	Extract & purify from gel	Store in -20°C
	Extract & purify AtoB4 from gel	Store in -20°C

GFP Brick Construction	o/n culture of AH044	MiniPrep

Week 10

Worked With	WorkFlow						July 20th - July 24th
Propane Plasmid 1 Construction	Gradient PCR for CAR1	Extract & purify from gel	Low yield	TOPO Blunt End cloning and transform into TOP10t	Screen with colony PCR	Screen more colonies
	PCR for new CAR	No product	Gradient PCR	PCR purification
	CAR3 PCR with DMSO run on gel	Extract purify from gel unclean product	TOPO Blunt End cloning and transform into TOP10t	Screen with colony PCR	Extract & purify from gel

Propane Plasmid 2 Construction	Run AtoB1 PCR with DMSO on gel	Extract & purify from gel	Low yield
	Run AtoB2 PCR with DMSO on gel	Extract & purify from gel	Low yield

Preparations Linearized Backbones	Transform AH043 into TOP10	o/n culture	MiniPrep	Restrict	Re-restrict

Week 11

Worked With	WorkFlow						July 27th - July 31st
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 1 Construction	Continue CAR1 colony screening	Screen more colonies	PCR purification
	PCR for new CAR2	Gradient PCR with KAPA polymerase	PCR purification
	Re-do colony PCR for CAR3 successful TOPO colony	Re-do colony PCR
		o/n culture of the colony	MiniPrep	PCR

Propane Plasmid 2 Construction	TOPO Blunt End Cloning for AtoB1 and transformation into TOP10	Screening with colony PCR	PCR purification

Cellulose Plasmid Construction	2nd PCR for BglX (from AH021)	Stored in -4°C over the weekend
	PCR for Cen1 & Cen2	Gradient PCR with KAPA polymerase	PCR purification for Cen2
			Gel purification for Cen1

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

Worked With	WorkFlow						August 3rd - August 8th
Preparations Linearized Backbones	Check restricted AH043 on gel	Use for CAR OE-PCR
	Re-restrict with high amount of DNA	Extract & purify from gel	Low yield
	New o/n culture of AH043 (needed more backbone)	MiniPrep	Remove insert	Use for CAR NEBuilder
Fixing Commercial Backbones	Restrict and ligate AH006 into AH031&AH033	Transform into TOP10

Propane Plasmid 1 Construction	OE-PCR without backbone	Weak product band	PCR purification	Gradient PCR	No product
	OE-PCR with backbone	Transform into TOP10	Red colonies
	NEBuilder protocol	Transform into TOP10	Screen with colony PCR	o/n culture	MiniPrep	Send for sequencing

Violacein Plasmid Construction	PCR purification for VioA 2nd PCR	Stored in -20°C

Cellulose Plasmid Construction	PCR purification for VioA 2nd PCR	Stored in -20°C

Amph protein	Gradient PCR P044&P045, P046&P051, P045&P046	Extract & purify from gel P044&P045	DNA not clean	Store in -20°C
		Stored P046&P051 and P045&P046 product gels to +4°C

GFP protein	Gradient PCR with P028&P029	PCR purification

Propane Measurement	o/n culture of Pauli's propane strain	Propane measurement	Results available next week

Week 13

Worked With	WorkFlow						August 10th - August 14th
Amphiphilic Protein Construction	PCR amph with P044&P045	No product	Gradient PCR	No product	dunnowhat

Preparations Backbones	Restrict AH043	Stored in -20°C

GFP & Amph Bricks Assembly	Restrict & ligate GFP and Amph into AH009	Transform into TOP10	Screen with colony PCR	o/n culture	MiniPrep	Check on gel	Send for sequencing

Propane Plasmid 2 Assembly	NEBuilder protocol	Transform into TOP10	Screen with colony PCR	Re-do NEBuilder and transfrom to TOP10	Screen with colony PCR	o/n culture of one potential colony	o/n culture turned red
	ELIC protocol	Transform into TOP10	No colonies

Cellulose Plasmid Assembly	NEBuilder protocol and transform into TOP10	Screen with colony PCR	Mistake in protocol	Re-do NEBuilder and transform into TOP10	Screen with colony PCR	o/n cultures and MiniPrep of white colonies	Restrict & check on gel
	ELIC protocol	Transform into TOP10	Screen with colony PCR	Mistake in protocol	Re-do ELIC and transform into TOP10	Screen with colony PCR

GFP Brick Validation	Restrict AH013, AH015 & AH007	Ligate AH007, AH013 & GFP	Transform into TOP10	RT over weekend
		Ligate AH007, AH015 & GFP	Transform into TOP10	RT over weekend

@@ Line 94: / Line 94: @@
          <li><a href="#week18" data-scroll="week18"><h4>Week 18</h4></a></li>
          <li><a href="#"><h4 style="border-top:solid;">To the Top</h4></a></li>
+        <li><a href="https://2015.igem.org/Team:Aalto-Helsinki/Lab" ><h4>To the Previous Page</h4></a></li>
        </ul>

Difference between revisions of "Team:Aalto-Helsinki/LabBook"

Revision as of 09:25, 28 August 2015

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

To the Top

To the Previous Page

Lab Book

Week 1

Worked With

WorkFlow

May 18th - May 22nd

Preparations Plates

Competent Cells

Propagations

Week 2

Worked With

WorkFlow

May 25th - May 29th

Preparations Propagating

Checking Competent Cells

Testing RBS's Assembly

Week 3

Worked With

WorkFlow

June 1st - June 5th

Preparations Propagating

Testing RBS's Assembly

Week 4

Worked With

WorkFlow

June 8th - June 12th

Preparations Propagating

Testing RBS's Assembly

Week 5

Worked With

WorkFlow

June 15th - June 19th

Testing RBS's Assembly

Preparations Linearized Backbones

Week 6

Worked With

WorkFlow

June 22nd - June 26th

Preparations Linearized Backbones

Testing RBS's Assembly

Propagation

Measurement

Week 7

Worked With

WorkFlow

June 29th - July 3rd

Preparations Linearized Backbones

Testing RBS's

Violacein Plasmid Construction

Week 8

Worked With

WorkFlow

July 6th - July 10th

Preparations Competent Cells

Cellulose Plasmid Construction

Testing RBS's

Violacein Plasmid Construction

Week 9

Worked With

May 18th -
May 22nd

Preparations
Plates

May 25th -
May 29th

Checking
Competent Cells

Testing RBS's
Assembly

June 1st -
June 5th

Preparations
Propagating

Testing RBS's
Assembly

June 8th -
June 12th

Preparations
Propagating

Testing RBS's
Assembly

June 15th -
June 19th

Testing RBS's
Assembly

Preparations
Linearized Backbones

June 22nd -
June 26th

Preparations
Linearized Backbones

Testing RBS's
Assembly

June 29th -
July 3rd

Preparations
Linearized Backbones

Violacein Plasmid
Construction

July 6th -
July 10th

Cellulose Plasmid
Construction

Violacein Plasmid
Construction

July 13th -
July 17th

Preparations
Propagations

July 20th -
July 24th

July 27th -
July 31st