Team:Groningen/Notebook/9 July 2009

From 2009.igem.org

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(Vectors)
(Vectors)
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|1,91
|1,91
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! MgCl
+
! pSBK3
| 24,4 ng/ul
| 24,4 ng/ul
-
|2,05 260/280
+
|2,05  
 +
 
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Revision as of 12:09, 17 August 2009

Igemhomelogo.png

Wet

GVP Cluster

Inoculation of TY-medium for Glycerol Stocks and Plasmid isolation

  • BBa_J23109 (labelled as no.1) vector BBa_J61002 in E.coli TOP10
  • BBa_J23100 (labelled as no.2) vector BBa_J61002 in E.coli TOP10 (red colony)
  • BBa_J23106 (labelled as no.3) vector BBa_J61002 in E.coli TOP10 (red colony)
  • The three plates showing new colonies were used to inoculate 5mL TY-medium with amp. (final conc. 100mg/ul).
  • The tubes were put in a waterbath at 37C (200 rpm) and grown o.n.

Transporters

First results!

Purification of HmtA (PA Q9I147 6xHis Tag)

  • SDS-gel electrophosis
  • Make a 12% SDS-gel
  • Fill gel with samples Wash 1-2 20mM imidazole, Wash1-2 40mM imidazole, Elute1-3 and ladders
  • Run 2h at 100V
  • Stain the gel in Coomassie blue (2h)
  • Destain with demiwater (2x, 10 min microwave)



Vectors

Measure concentration with Nanodrop

Component concentrations ng/ul 260/280
pSB1AC3 205,7 1,91
pSBK3 24,4 ng/ul 2,05

Dry

A lot of time was spent on finding dissociation constants and figuring out how ArsR/D work, and thinking of how to model their interaction. An input function was derived for the ArsR/D operator, but it is not yet completely clear how this should be used in a model (it is a function of the ArsR/D concentration, but at the same time produces ArsR/ArsD). It would probably be better to start with As(III) and derive an input function from that, if at all possible.


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