Team:Groningen/Notebook/28 August 2009

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Wet

GVP Cluster

DONE Isolate plasmids from o.n. cultures
TODO Restriction analysis of plasmids for correct ligation of GVP behind LAC/pBAD promoter
TODO Test control of bouyancy in Saline solution
TODO Order synthetic DNA for GVP
TODO Test promoter strenght compared to BBa_J23101 promoter (Sven)
TODO Enter sequences of constructs to Sandbox


Overnight Precultures

All tubes showed growth of E.coli cells, and must have ampicillin resistance on a plasmid(The tubes differed in cell density though). To be sure of correct ligation products in the plasmids an isolation will be performed, followed by restriction analysis. If positive, the plasmids will be sent for sequencing.

Overnight Plates

The plates grown overnight for glycerol stocks showed single colony growth on the second and third wipe stripes. The plates were stored in the fridge for preculture inoculation on monday (31/8).

→ The colonies for pArsR+RBS showed red colonies, and the pCueO+RBS/pZntR+RBS showed normal yellow/white colonies.

Restriction for Assembly

The vector pSB1AC3 containing the LAC/pBAD inducible promoters and GVP were cut with PstI and EcoRI to check correct ligation product.

  • 5μL plasmid in MQ
  • 11μL MQ (end volume of 20μL)
  • 2μL Fast digest buffer
  • 1μL PstI fast digest enzyme
  • 1μL EcoRI fast digest enzyme

Restriction was kept at 37C for 30 min., followed by addition of 6x loading dye and put on ice until used for gel analysis.

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