Team:Groningen/Notebook/11 August 2009

From 2009.igem.org

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'''Plates'''
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LB-agar plates with 50 μg/mL ampicillin were freshly prepared, and one drop of each o.n. preculture was spotted and subsequently swiped three times with sterile needle. The plates were stored at 37°C overnight.
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'''Precultures'''
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Over night cultures in 5mL LB-amp<sub>50</sub> medium were prepared from the following colonies:
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→ E.coli TOP10 pSB1AC3-med. const. promoter - GVP (amp.) (4x) (1 to 4)
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→ E.coli TOP10 pSB1AC3-low const. promoter - GVP (amp.) (4x) (5 to 8)
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and put in the 37°C waterbath at 200 rpm.
===Transporters===
===Transporters===

Revision as of 09:30, 11 August 2009

Igemhomelogo.png

Wet

GVP Cluster

DONE isolate plasmids from overnight precultures
DONE plate on cultures for storage in fridge (amp)
TODO cut plasmid and GVP containing plasmid for ligation (ligation tomorrow)
TODO purify wanted fragments (tomorrow)
DONE make o.n. precultures from plate colonies
www.sigmaaldrich.com

Plasmid Purification

Plasmid isolation was performed on the cultures of E.coli TOP10 containing plasmids J61002 with high, medium and low constitutive promoters 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 20μL MQ and stored in the fridge

Concentrations

Plasmid Conc. ng/μL 260/280 260/230 -20 box (michael Restriction Control
pJ61002-BBa_J23100 (high) 174.6 1.94 2.40 E-8 No
pJ61002-BBa_J23106 (medium) 149.2 1.95 2.41 E-9 No
pJ61002-BBa_J23109 (low) 152.8 1.94 2.38 E-10 No

Plates

LB-agar plates with 50 μg/mL ampicillin were freshly prepared, and one drop of each o.n. preculture was spotted and subsequently swiped three times with sterile needle. The plates were stored at 37°C overnight.

Precultures

Over night cultures in 5mL LB-amp50 medium were prepared from the following colonies:

→ E.coli TOP10 pSB1AC3-med. const. promoter - GVP (amp.) (4x) (1 to 4) → E.coli TOP10 pSB1AC3-low const. promoter - GVP (amp.) (4x) (5 to 8)

and put in the 37°C waterbath at 200 rpm.

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