Team:Groningen/Notebook/17 August 2009

From 2009.igem.org

(Difference between revisions)
(New page: {{Team:Groningen/Notebook/Day/Header}} ==Wet== ===GVP Cluster=== :→ {{todo}} Restriction for Assembly :→ {{todo}} Gel purification of plasmid :→ {{todo}} Ligation of metal promote...)
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:→ {{todo}} Transformation of E.coli TOP10 cells
:→ {{todo}} Transformation of E.coli TOP10 cells
:→ {{todo}} Test promoter/GVP constructs (grow precultures)
:→ {{todo}} Test promoter/GVP constructs (grow precultures)
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:→ {{todo}} Grow o.n. cultures to check ligation of pSB3K3-H/L-GVP constructs of last week
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'''Tranformation'''
'''Tranformation'''
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===Transporters===
===Transporters===

Revision as of 07:05, 17 August 2009

Igemhomelogo.png

Wet

GVP Cluster

TODO Restriction for Assembly
TODO Gel purification of plasmid
TODO Ligation of metal promoter oligo's into vector BBa_J61002
TODO Transformation of E.coli TOP10 cells
TODO Test promoter/GVP constructs (grow precultures)
TODO Grow o.n. cultures to check ligation of pSB3K3-H/L-GVP constructs of last week


Restriction for Assembly

The vector BBa_J61002 containing the high constitutive promoter was cut with EcoRI and SpeI to create correct ends for insert of metal promoter oligo's.

  • 8 μL plasmid in MQ (1.0μg)
  • 8 μL MQ (end volume of 20μL)
  • 2μL Fast digest buffer
  • 1μL PstI fast digest enzyme
  • 1μL SpeI fast digest enzyme

Restriction was kept at 37C for 30 min. and put on ice until used for gel purification.

Purification

Concentrations

Plasmid Conc. ng/μL 260/280 260/230 -20 box (michael Restriction Control
BBa_J61002-H SpeI/EcoRI ? ? ? ? ?

Ligation


Tranformation

Transporters

Metal Accumulation

Vectors

Dry

April
MTWTFSS
    1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30
May
MTWTFSS
        1 2 3
4 5 6 7 8 9 10
11 12 13 14 15 16 17
18 19 20 21 22 23 24
25 26 27 28 29 30 31
June
MTWTFSS
1 2 3 4 5 6 7
8 9 10 11 12 13 14
15 16 17 18 19 20 21
22 23 24 25 26 27 28
29 30
July
MTWTFSS
    1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30 31
August
MTWTFSS
          1 2
3 4 5 6 7 8 9
10 11 12 13 14 15 16
17 18 19 20 21 22 23
24 25 26 27 28 29 30
31
September
MTWTFSS
  1 2 3 4 5 6
7 8 9 10 11 12 13
14 15 16 17 18 19 20
21 22 23 24 25 26 27
28 29 30
October
MTWTFSS
      1 2 3 4
5 6 7 8 9 10 11
12 13 14 15 16 17 18
19 20 21 22 23 24 25
26 27 28 29 30 31
November
MTWTFSS
            1
2 3 4 5 6 7 8
9 10 11 12 13 14 15
16 17 18 19 20 21 22
23 24 25 26 27 28 29
30