Team:Groningen/Notebook/4 September 2009

From 2009.igem.org

(Difference between revisions)
(GVP Cluster)
(GVP Cluster)
Line 10: Line 10:
'''O.n. precultures'''
'''O.n. precultures'''
-
:→ All twelve o.n. precultures showed bacterial growth, and could be used to isolate plasmid. The first four are registry vector pSB2K3 with RFP as reporter (plate 1, 7C) which were transformed on tuesday. The isolated plasmid is used to house all GVP constructs to combine with the vector pSB1AC3 with transporter and accumulation genes.
+
:→ All four o.n. precultures showed bacterial growth, and could be used to isolate plasmid. The four are registry vector pSB1A2 with pLacI/pBad-araC and GVP. The isolated plasmid is used for transformation into pSB2K3 and sent for sequencing.
-
:→ The next eight tubes contain our own designed biobricks and isolated plasmid will be sent to the registry on friday!
 
[[Image:Plasmid Miniprep Kit Flow Protocol.gif|thumb|150px| www.sigmaaldrich.com]]
[[Image:Plasmid Miniprep Kit Flow Protocol.gif|thumb|150px| www.sigmaaldrich.com]]
Line 34: Line 33:
|'''Restriction Control'''
|'''Restriction Control'''
|-
|-
-
|pSB2K3 (7C) no.1
+
|pSB1A2 pLacI-GVP no.1
-
|178.1
+
|405.3
-
|1.87
+
|1.85
-
|2.32
+
|2.36
-
|D-1
+
|D-6
 +
|Yes (EcoRI/PstI)
 +
|-
 +
|pSB1A2 pLacI-GVP no.2
 +
|34.0
 +
|2.01
 +
|2.20
 +
|D-7
 +
|Yes (EcoRI/PstI)
 +
|-
 +
|pSB1A2 pBad/araC-GVP no.1
 +
|27.5
 +
|2.09
 +
|2.16
 +
|D-8
 +
|Yes (EcoRI/PstI)
 +
|-
 +
|pSB1A2 pBad/araC-GVP no.2
 +
|33.3
 +
|1.96
 +
|2.06
 +
|D-9
|Yes (EcoRI/PstI)
|Yes (EcoRI/PstI)
|}
|}

Revision as of 07:51, 4 September 2009

Igemhomelogo.png

Wet

GVP Cluster

BBa_F2620


O.n. precultures

→ All four o.n. precultures showed bacterial growth, and could be used to isolate plasmid. The four are registry vector pSB1A2 with pLacI/pBad-araC and GVP. The isolated plasmid is used for transformation into pSB2K3 and sent for sequencing.


www.sigmaaldrich.com

Plasmid Purification

Plasmid isolation was performed on the cultures of E.coli TOP10 containing the above mentioned plasmids with the "Sygma-Aldrich™ GenElute™ Plasmid Miniprep Kit".

  • From each tube 4mL of culture was collected in a 2.0mL cup, and the cells were pelleted by centrifugation for 1 min. at max. speed and the supernatant discarded.
  • Plasmids were eluted with 30μL MQ and stored in the fridge


Concentrations

Plasmid Conc. ng/μL 260/280 260/230 -20 box (michael Restriction Control
pSB1A2 pLacI-GVP no.1 405.3 1.85 2.36 D-6 Yes (EcoRI/PstI)
pSB1A2 pLacI-GVP no.2 34.0 2.01 2.20 D-7 Yes (EcoRI/PstI)
pSB1A2 pBad/araC-GVP no.1 27.5 2.09 2.16 D-8 Yes (EcoRI/PstI)
pSB1A2 pBad/araC-GVP no.2 33.3 1.96 2.06 D-9 Yes (EcoRI/PstI)

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