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             <p>PCR<br/> purification</p>
 
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Revision as of 13:19, 11 August 2015

Lab Book

An example of what the table could look like:

Construct
worked with

Things done

Lab 1.1.2015
Monday

CAR

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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

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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

Plates

Making Cm
plates

Stored in
+4°C

Competent Cells

Preparing
solutions for
competent cells

Stored in
+4°C

Competent Cells

o/n culture
of E. coli
TOP10

Chemical
competent
cells protocol

Tranformation
efficiency kit

Competent Cells

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

Construct
worked with

Things done

May 25th - May 29th

Propagations

Transformed
AH007-AH009,
AH005 & AH019
into TOP10

AH007-AH009
& AH019

o/n cultures

MiniPrep

No colonies
for AH005

Re-transform
into TOP10

Plates in rt
over weekend

Competent Cells

o/n culture
of BL21(DE3)
transformants

MiniPrep

PCR with
VF2 & VR

  Smeared gels

Assembly
Testing RBS's

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

AH001+
   AH003+AH008
-> AH013

Repeat
ligation

Restrict &
extract
from gel

 Transformation
into TOP10

Plates in rt
over weekend

Propagations

o/n culture of
  AH001-AH004
& AH006

MiniPrep

Week 3

Construct
worked with

Things done

June 1st - June 5th

Assembly
Testing RBS's

No colonies
for AH011
or AH014

re-assemble

Restrict &
extract
from gel

Ligate &
tranform
into TOP10

No colonies

Assembly
Testing RBS's

AH005 &
AH013 OK

o/n cultures

MiniPrep

Propagations

Transform
    AH020 & AH021
into TOP10

Plates in rt
 over weekend

Week 4

Construct
worked with

Things done

June 8th - June 12th

Assembly
Propagations

o/n cultures
for AH020
& AH021

MiniPrep

Assembly
Testing RBS's

Re-assembling
   AH011, AH013
& AH014

Transform into TOP10

o/n cultures

MinPrep

Assembly
Testing RBS's

Assembling
AH015

  Ligate AH002+
  AH005+AH008

Restrict &
extract
from gel

Transform into TOP10

o/n cultures

MiniPrep

Propagations

Transform
    AH020 & AH021
into TOP10

Plates in rt
over weekend

Week 5

Week 6

Construct
worked with

Things done

Week 6

Assembly
Testing RBS's

Re-assembling AH016 &
AH018

Transformation
into BL21(DE3)

No colonies

Testing RBS's

o/n cultures of
  AH016 & AH018
in TOP10

MiniPrep

Transform
into BL21(DE3)

Plates in rt
over the
weekend

Measurement
Testing RBS's

AH018 in
BL21(DE3)

o/n culture

IPTG induction &
    blue chromoprotein
measurement

No blue chromoprotein

Refresh on IPTG plate

No blue chromoprotein

Producing linearized backbones

o/n cultures of
AH007-AH009

MiniPrep

Restrict &
extract
from gel

Week 7

Construct
worked with

Things done

Week 6

Preparations
Testing RBS's

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

Measurement
Testing RBS's

Refresh AH016 &
AH018 on
IPTG plate

No blue
chromoprotein

Violacein construct

PCR AH20 with
P012&P013

PCR
purification

VioA waits
for 2nd PCR

PCR AH20 with
P024&P025

PCR
unsuccesful

PCR AH20 with
P035&P025

PCR
purification

VioE waits
for 2nd PCR

Week 8

Construct
worked with

Things done

Lab 1.1.2015
Monday

Constructing Cellulose system

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

Preparations

Preparing
  PK BL21(DE3)
competent cells

Cells stored
in -80°C

Testing RBS's

Transform
AH016 & AH018
into PK BL21(DE3)

Refresh
transformants
on IPTG plates

No blue
chromoprotein

Violacein Construct

Gradient PCR for
AH020 with
P024&P025

PCR
purification

Week 9

Week 10

Week 11

Week 12

Week 13

Week 14

Week 15

Week 16

Week 17

Week 18