Team:Groningen/Notebook/31 July 2009

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Revision as of 10:01, 31 July 2009 by JolandaWitteveen (Talk | contribs)

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Wet

GVP Cluster

Transporters

GlpF

  • Restiction/ligase and transformation of GlpF in psB1AC3

Restiction

10ul GlpF PCR
2 ul Buffer
05.ul EcoRI
0.5 ul SpeI
7 uL MQ
- 37 30min
-Inactivation on Gel
-Extraction from gel

Metal Accumulation

ArsR-MBP fusion

  • colony PCR MBP:
colony PCR MBP
Component amount
2x Phusion MM 12.5 uL
MQ 9.5 uL
MBP Fw 1 uL
MBP Rev 1 uL
Colony MBP 1 uL
Touchdown MBP program Temperature Time
Denaturing 98° 2.00 min
Touchdown 10X 60->50
Denaturing 98° 30 sec
Annealing 60°->50° 30 sec
Elongation 72° 30 sec
End cycles
Start Cycles 25X
Denaturing 98° 30sec
Annealing 60° 30 sec
Elongation 72° 30 sec
End cycles
Final elongation 72° 7 min
Hold Forever
  • Overnight culture MBP

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