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On the following pages you can find recipes for standard
culture media which are generally used for growing Fusarium species
and for standard molecular biological procedures
We normally make sterile stock solutions of the individual
components and then mix them under sterile conditions. For solid media we
typically start with a water agar solution which has been melted and then cooled
to 55 oC. Sterilization: autoclavation (A) or sterile filtration (SF)
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CAS |
100 ml |
500 ml |
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2.5x Salt solution (SF or A) |
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40.0 ml |
200.0 ml |
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20 % (w/v)
Glucose
(SF or A) |
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0.90 ml |
4.5 ml |
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20 %
(v/v)
Glycerol (SF or A) |
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2.50 ml |
12.5 ml |
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(Agar)
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(2
g) |
(10.0 g) |
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H2O (A) |
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fill to 94 ml |
fill to 470 ml |
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MES-solution
(1 M) (SF) |
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4.0 ml |
20.0 ml |
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Acetosyringon (10 mM) (SF)
(FRESH!!) |
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2.0 ml |
10.0 ml |
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2.5X Salt solution (1 l): |
CAS |
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KH2PO4 |
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3.625 g |
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K2HPO4 |
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5.125 g |
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NaCl |
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0.375 g |
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MgSO4, 7H2O |
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1.250 g |
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CaCl2, 2H2O
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0.165 g |
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FeSO4, 7H2O |
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0.0062 g |
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(NH4)2SO4
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1.250 g |
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Add MilliQ water to |
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1000 ml |
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Dissolve each salt ONE AT A TIME |
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Filter-sterilize (0.2
mm filter) |
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Store at room temperature. |
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10 mM Acetosyringone (AS) |
CAS |
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Acetosyringone |
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0.01962 g |
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MilliQ-water |
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10 ml |
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Adjust the pH to 8 with 5 M KOH
(7.013
g KOH in 25 ml water) |
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Filter-sterilize (0.2
mm filter) |
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Store
in 10 ml aliquots at -20 oC |
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1 M MES
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CAS |
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MES
hydrate |
4432-31-9 |
19.52 g |
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MilliQ-water |
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80 ml |
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Adjust pH to 5.3 with 5 M KOH. |
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Bring the volume to 100 ml with
MilliQ-water |
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Filter-sterilize (0.2
mm filter) |
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Store
in 20 ml aliquots at -20 oC |
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back to the top
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CAS |
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500 ml |
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20 % Glucose
(SF) |
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31.25 ml |
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50 mM Aspargin
(SF) |
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100.0 ml |
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0.21
M MgSO4
(26 g/ 500 ml) (A) |
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5.0 ml |
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1.12
M KH2PO4 + 0.7 M KCl
(pH 6) (A) |
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5.0 ml
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1000
x Trace elements (A) |
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0.5 ml |
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Agar |
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10.0 g |
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MilliQ |
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348.3 ml |
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1000 x Trace elements for DFM |
CAS |
1000 ml |
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Na2B4O7 * 10H2O,
(Borax = Natriumtetraborat) |
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40 mg |
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CuSO4 * 5H2O |
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400 mg |
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FeSO4 * 7H2O |
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1,2 g |
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MnSO4 * 1H2O |
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700 mg |
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NaMoO2
* 2H2O |
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800 mg |
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ZnSO4 * 7H2O |
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10 g |
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Add MilliQ water to |
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1000 ml |
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Dissolve each salt ONE AT A TIME! |
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Autoclave
the solution (note the the iron will oxidate and turn the solution red) |
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back to the top
Bells (Bell,
A.A. Pest Manag Sci 2003)
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CAS |
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1000 ml |
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(- all components are mixed from
salts) |
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Maltose |
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20 g |
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Urea |
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0.140
g |
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KH2PO4 |
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1 g |
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MgSO4
*6H2O |
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0.5
g |
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KCl |
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0.5
g |
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CaCO3 |
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0,5 g |
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MES |
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1 ml |
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MilliQ
water |
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fill to 990
ml |
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Adjust pH to 5.1 with 1 M
K2HPO4 |
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Add water to 1000 ml and autoclave
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MES = Minor Elements Solution
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CAS |
1000 ml |
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FeSO4 *7H2O |
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10 g |
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ZnSO4
*7H2O |
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50 g |
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CuSO4
*5H2O |
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2.5
g |
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MnSO4 *H2O |
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0.5
g |
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NaB4O7 |
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0.5
g |
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NaMoO4
*2H2O |
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0.5
g |
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Citric acid |
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50 g |
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Add MilliQ water to |
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1000 ml |
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Dissolve each salt ONE AT A TIME! |
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Autoclave
the solution (note the the iron will oxidate and turn the solution red) |
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back to the top
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CAS |
1000
ml |
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(- all components are mixed from
salts) |
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Yeast extract |
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20 g |
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Sucrose |
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150 g |
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MgSO4
*7H2O |
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0.5 g |
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Agar |
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20 g |
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Trace solution |
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1.0 ml |
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MilliQ
water |
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fill to 990
ml |
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Adjust pH to 6.5 |
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Add water to 1000 ml and autoclave
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Trace solution |
CAS |
100 ml |
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ZnSO4
* 7 H2O |
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10 g |
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CuSO4
* 5 H2O |
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50 g |
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MilliQ
water |
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fill to 100 ml |
back to the top
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Chop 400 g baby carrots in
food processor |
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Put carrot in 2
litre glass beaker, add 400 ml deionised water, cover loosely with Saran
wrap and microwave 2 x 10 min. |
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Let cool 1 – 2 hrs |
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Puree in food processor |
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Return to beaker, add 500
ml water, mix well |
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Dispense evenly into 5 x
500 ml Pyrex bottles (ca. 225 ml per bottle) containing 4 g agar per bottle
(~ 1.8 % agar) |
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Autoclave 20 min |
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back to the top
Fries medium
(Pringle and Braun (1957), Phytopathology (47) p. 369-371)
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CAS |
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1000
ml |
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Ammonium tartrate
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5.0 g |
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NH4NO3 |
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1.0 g |
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KH2PO4 |
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1.0 g |
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MgSO4
*6H2O |
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0.46 g |
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NaCl |
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0.1 g |
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CaCl2
*2 H2O |
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0.13 g |
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Sucrose |
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30.0 g |
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MnSO4
*H2O |
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0.76 mg |
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Boric acid |
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1.0 mg |
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CuSO4
*5H2O |
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0.36 mg |
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ZnSO4
*7H2O |
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0.1 mg |
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FeSO4
*7H2O |
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20 mg |
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Bacto Agar |
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20 g |
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MilliQ water |
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fill to 1000 ml |
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Measured pH in final media it should
be 5.6 |
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Autoclavate |
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back to the top
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1000
ml |
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Yeast extract |
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10 g |
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Peptone |
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20 g |
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MilliQ water |
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Fill to 950 ml |
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Autoclave |
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Add 50 ml 50 % glucose (SF) before use
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back to the top
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1000
ml |
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Succrose |
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30 g |
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KH2PO4
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1 g |
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MgSO4 x 7 H2O |
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0,5 g |
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KCl |
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0,5 g |
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TES** |
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0,2 ml |
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NaNO3
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2 g |
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Bacto Casamino |
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2,5 g |
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Yeast extract |
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1 g |
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Vitamin solution |
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10 ml |
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MilliQ water |
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fill to 1000 ml |
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Vitamin solution |
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Thiamine HCl |
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100 mg |
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Riboflavin |
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30 mg |
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Pyridoxine HCl |
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75 mg |
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D-pantothenate Ca |
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200 mg |
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P-aminobenzoic acid |
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5 mg |
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Nicotinamide
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75 mg |
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Choline Cl |
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200 mg |
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Folic acid |
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5 mg |
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D-biotin |
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5 mg |
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Myo-inositol |
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4 g |
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50% ethanol |
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fill to 1000 ml |
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TES** |
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Citric
acid |
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5 g |
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Zinc sulfate x H2O |
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5 g |
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Fe(NH4)2(SO4)2
x H2O |
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1 g |
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CuSO4 x H2O |
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0,25 g |
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MnSO4 x H2O |
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50 mg |
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H3BO4 |
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50 mg |
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NaMoO4 x H2O |
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50
mg |
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MilliQ water |
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95 ml |
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back to the top
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1000
ml |
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Succinic acid (C4H4O4Na2 * 6H2O) |
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50 g |
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NaNO3 |
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12.1 g |
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Glucose |
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1 g |
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Vogels salte 50x (-N, -C) |
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20 ml |
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MQ water |
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fill to
1000 ml |
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50 x Vogels stalte
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Na3-citrat * 2H2O |
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125 g |
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KH2PO4 |
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250 g |
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MgSO4 * 7H2O |
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10 g |
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CaCl2 *2H2O |
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5 g |
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200x Trace opløsning |
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5 ml |
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MQ water |
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fill to 1000 m |
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200 x Trace |
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Citric acid
*H2O |
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5 g |
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ZnSO4 *7H2O |
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5 g |
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Fe(NH4)2(SO4)2
*6H2O |
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1 g |
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CuSO4 *5H2O |
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0.25
g |
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MgSO4 *H2O |
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0.05
g |
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H3BO3 |
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0.05
g |
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Na2MoO4 *
2H2O |
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0.05
g |
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MQ water |
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fill to 1000 ml |
back to the top
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1000
ml |
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Bacto-tryptone |
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10 g |
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Bacto-yeast
extract |
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5 g |
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NaCl (solid) |
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10 g |
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MilliQ
water |
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add to 950 ml |
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Adjust pH with 5 M NaOH |
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E. coli: 7.4 to 7.5 |
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A. tumefaciens: 7.7 - 7.8 |
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MilliQ |
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add to 1000 ml |
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For solid medium add 10 g/L of agar |
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back to the top
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