Team:Aalto-Helsinki/LabBook
Worked With |
WorkFlow |
May 18th - |
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Preparations |
Making Cm |
Stored in |
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Competent Cells |
Preparing |
Stored in |
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o/n culture |
Chemical |
Tranformation |
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o/n culture |
Chemical |
Tranformation |
Colonies are |
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Propagations |
Transformed |
AH001-AH004 |
Stored in |
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No colonies for |
Click here to read the full lab notes of week 1.
Worked With |
WorkFlow |
May 25th - |
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Preparations Propagating |
Transformed |
AH007-AH009 |
o/n cultures |
MiniPrep |
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o/n culture of |
MiniPrep |
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No colonies |
Re-transform |
Plates in rt |
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Checking |
o/n culture |
MiniPrep |
PCR with |
Smeared gels |
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AH001+ |
Repeat |
Restrict & |
Transform |
Plates in |
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Testing RBS's |
Restrict |
Ligation |
AH002+ |
Repeat |
Restrict & |
Transform |
Plates in |
AH001+ |
Repeat |
Restrict & |
Transform |
Plates in |
Click here to read the full lab notes of week 2.
Worked With |
WorkFlow |
June 1st - |
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Preparations |
Transform |
Plates in rt |
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Testing RBS's |
No colonies |
re-assemble |
Restrict & |
Ligate & |
No colonies |
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AH005 & |
o/n cultures |
MiniPrep |
Assembling |
Ligate AH002+ |
Restrict & |
Transform into TOP10 |
o/n cultures |
No colonies |
Click here to read the full lab notes of week 3.
Worked With |
WorkFlow |
June 8th - |
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Preparations |
o/n cultures |
MiniPrep |
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Testing RBS's |
Re-assembling |
Transform into TOP10 |
o/n cultures |
MinPrep |
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Assembling |
Ligate AH002+ |
Restrict & |
Transform into TOP10 |
o/n cultures |
MiniPrep |
Click here to read the full lab notes of week 4.
Worked With |
WorkFlow |
June 15th - |
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AH016 |
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Colonies in |
o/n cultures |
AH018 OK |
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Testing RBS's |
Screening AH011 |
AH011, AH013 |
Assembling |
Transform into TOP10 & BL21(DE3) |
T7 promoter |
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AH014 |
Assembling AH017 |
Restrict & |
Transform into BL21(DE3) |
Colonies are |
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Re-assembling |
Restrict |
Transform into |
Colonies are |
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Preparations |
o/n cultures of |
MiniPrep |
Restrict & |
Low |
Click here to read the full lab notes of week 5.
Worked With |
WorkFlow |
June 22nd - |
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Preparations |
o/n cultures of |
MiniPrep |
Restrict & |
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Testing RBS's |
Re-assembling AH016 & |
Transform |
No colonies |
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Propagation |
o/n cultures of |
MiniPrep |
Transform |
Plates in rt |
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Measurement |
AH018 in |
o/n culture |
IPTG induction & |
No blue chromoprotein |
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Refresh on |
No blue chromoprotein |
Click here to read the full lab notes of week 6.
Worked With |
WorkFlow |
June 29th - |
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Preparations |
Transform |
No colonies |
Use iGEM's |
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Testing RBS's |
o/n cultures of |
IPTG |
No Blue |
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Refresh AH016 & |
No blue |
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Violacein Plasmid |
PCR AH20 |
PCR |
VioA waits |
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PCR AH20 |
PCR |
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PCR AH20 |
PCR |
VioE waits |
Click here to read the full lab notes of week 7.
Worked With |
WorkFlow |
July 6th - |
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Preparations Competent Cells |
Preparing |
Cells stored |
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Cellulose Plasmid |
PCR AH021 |
Problem |
Re-do PCR |
PCR |
BglX waits for |
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AH029 into |
Transformants |
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Testing RBS's |
Transform AH016 |
Refresh |
No blue |
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Violacein Plasmid |
Gradient PCR for |
PCR |
Click here to read the full lab notes of week 8.
Worked With |
WorkFlow |
July 13th - |
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Preparations |
o/n culture |
MiniPrep |
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CAR2 OK |
PCR |
An illegal |
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Propane Plasmid 1 Construction |
PCR for |
CAR1 & 3 |
Gradient PCR |
PCR with |
Store |
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PCR with DMSO |
No product |
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PCR with |
Store |
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Propane Plasmid 2 Construction |
PCR for |
Gradient PCR |
PCR with |
Store |
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Gradient PCR |
Extract & |
Store |
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Extract & |
Store |
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GFP Brick Construction |
o/n culture |
MiniPrep |
Click here to read the full lab notes of week 9.
Worked With |
WorkFlow |
July 20th - |
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Propane Plasmid 1 Construction |
Gradient PCR |
Extract & |
Low yield |
TOPO Blunt End |
Screen with |
Screen more |
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PCR for new CAR |
No product |
Gradient PCR |
PCR purification |
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CAR3 PCR |
Extract |
TOPO Blunt End |
Screen with |
Extract & |
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Propane Plasmid 2 Construction |
Run AtoB1 |
Extract & |
Low yield |
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Run AtoB2 |
Extract & |
Low yield |
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Preparations Linearized Backbones |
Transform AH043 into TOP10 |
o/n culture |
MiniPrep |
Restrict |
Re-restrict |
Click here to read the full lab notes of week 10.
Worked With |
WorkFlow |
July 27th - |
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Preparations Linearized Backbones |
Re-restrict |
Gel purification |
Low yield |
Re-restrict |
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Propagations |
o/n culture |
MiniPrep |
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Transform |
o/n culture |
MiniPrep |
Low yield |
o/n culture |
MiniPrep |
Low yield, |
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Propane Plasmid 1 Construction |
Continue |
Screen more |
PCR |
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PCR for |
Gradient PCR |
PCR |
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Re-do colony |
Re-do |
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o/n culture of |
MiniPrep |
PCR |
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Propane Plasmid 2 Construction |
TOPO Blunt End |
Screening |
PCR |
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Cellulose Plasmid Construction |
2nd PCR |
Stored in -4°C |
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PCR for |
Gradient PCR |
PCR |
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Gel purification |
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Violacein Plasmid Construction |
2nd PCR |
Stored in -4°C |
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KAPA gradient |
Gel purification |
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PCR |
Worked With |
WorkFlow |
August 3rd - |
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Preparations Linearized Backbones |
Check |
Use for CAR |
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Re-restrict |
Extract & |
Low yield |
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New o/n culture |
MiniPrep |
Remove insert |
Use for CAR |
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Fixing Commercial Backbones |
Restrict and |
Transform into TOP10 |
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Propane Plasmid 1 Construction |
OE-PCR |
Weak product |
PCR |
Gradient |
No product |
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OE-PCR |
Transform |
Red colonies |
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NEBuilder |
Transform |
Screen with |
o/n culture |
MiniPrep |
Send for |
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Violacein Plasmid Construction |
PCR purification |
Stored |
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Cellulose Plasmid Construction |
PCR purification |
Stored |
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Amph protein |
Gradient PCR |
Extract & |
DNA |
Store |
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Stored P046&P051 |
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GFP protein |
Gradient PCR |
PCR |
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Propane Measurement |
o/n culture |
Propane measurement |
Results |
Worked With |
WorkFlow |
August 10th - |
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Amphiphilic Protein Construction |
PCR amph |
No product |
Gradient |
No product |
dunnowhat |
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Preparations Backbones |
Restrict AH043 |
Stored |
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GFP & Amph Bricks |
Restrict & |
Transform |
Screen with |
o/n culture |
MiniPrep |
Check on |
Send for |
Propane Plasmid 2 Assembly |
NEBuilder |
Transform |
Screen with |
Re-do NEBuilder |
Screen with |
o/n culture of |
o/n culture |
ELIC protocol |
Transform |
No colonies |
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Cellulose Plasmid Assembly |
NEBuilder protocol |
Screen with |
Mistake in |
Re-do NEBuilder |
Screen with |
o/n cultures and |
Restrict & |
ELIC |
Transform |
Screen with |
Mistake in |
Re-do ELIC |
Screen with |
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GFP Brick Validation |
Restrict AH013, |
Ligate AH007, |
Transform |
RT over |
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Ligate AH007, |
Transform |
RT over |
Worked With |
WorkFlow |
August 17th - |
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Preparations |
Transform |
o/n culture |
MiniPrep |
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Propane Plasmid 1 Assembly |
Restrict & |
Transform into TOP10 |
o/n culture |
MiniPrep |
Restrict & |
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Sequencing of |
Send construct in AH009 for sequencing |
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Validation |
Transform into |
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Amphiphilic Propane 1 |
Gradient PCR |
CAR amph3 |
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CAR amph4 |
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Propane Plasmid 2 |
OE-PCR |
No product |
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Check old |
Re-transform |
o/n cultures of |
MiniPrep |
Restrict & |
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Cellulose Plasmid Construction |
OE-PCR |
Extract & |
Concentration too low |
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PCR |
NEBuilder with AH009 |
Transform into |
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Check old |
Re-transform into TOP10 |
o/n of colorless |
MiniPrep |
Restrict & |
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GFP Assembly |
2A assembly for |
Transform |
no colonies |
3A assembly |
Transform |
No colonies |
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Propagate GFP in AH009 |
MiniPrep |
Send to Slovenia |
Worked With |
WorkFlow |
August 17th - |
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Propane Plasmid 1 |
Prepare |
Unclear |
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Analytical |
Wrong sized |
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Prepared |
Stored |
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Propane Plasmid 2 |
Repeat |
Transform into TOP10 |
Colony PCR |
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Repeat |
Transform into TOP10 |
Colony PCR |
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GFP Biobrick Assembly |
2A Assemble |
Transform |
O/n cultues |
MiniPrep |
Restrict & |
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3A Assembly |
Transform |
dunnowhat |
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GFP+Amph Assembly |
3A Assemble |
Transform |
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Cellulose Plasmid Construction |
Repeat |
Transform into TOP10 |
Colony PCR |
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Repeat |
Transform into TOP10 |
Colony PCR |
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Amphiphilic Propane1 |
NEBuilder |
Transform |
Colony PCR |
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O/n culture |
MiniPrep |