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Revision as of 14:10, 24 August 2010 (view source) Joelkuiper (Talk | contribs) (New page: -Make plates with a thick layer of medium, 100 ml medium per big Petri dish.Medium of Streptomyces is mannitol soya flour medium (MS or SFM) 300 ml per 1L bottle to avoid boiling over in a...) ← Older edit		Latest revision as of 17:19, 9 September 2010 (view source) Neima (Talk | contribs)
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-	-Make plates with a thick layer of medium, 100 ml medium per big Petri dish.Medium of Streptomyces is mannitol soya flour medium (MS or SFM) 300 ml per 1L bottle to avoid boiling over in autoclave
-	*Agar                          6 g	+	'''Concentrations for antibiotics '''
-	*D-mannitol                    6 g	+
-	*Soya flour                    6 g	+
-	*Demiwater                      300ml	+
-	*Plating ΔrdeAB spores          10^5 /plate	+
-	-Incubate for 7 days at 30 °C	+	Antibiotic                 Stock                         E.coli                         B.subtilis
-		+	*Ampicillin (Amp) (in H2O)   50 mg/ml (4 °C) 100(μg/ml) (:500)
-	-Harvest mycelium with razorblade	+	*Kanamycin (Km)           10 mg/ml (4 °C) 20 (μg/ml) (:500)                        10 (μg/ml) (:100)
		+	*Spectinomycin (Spec) (in H2O)   10 mg/ml (4 °C) 100(μg/ml) (:100)                 100 (μg/ml) (:100)
		+	*Erythromycin (Ery) E.coli stock  10 mg/ml (-18 °C) 150 (μg/ml) (:66.6)
		+	*Erythromycin(Ery) B.subtilis stock 1 mg/ml (-18 °C)                                 2-5 (μg/ml)
		+	*Tetracyclin (Tet) (50% EtOH)     3 mg/ml (-18 °C) 12 (μg/ml) (:250)                 6(μg/ml) (:500)
		+	*Chloramphenicol (Cm) (50% EtOH)    5 mg/ml (-18 °C) 10-15 (μg/ml) (:500/333)         5(μg/ml) (:1000)
		+	*Lyncomycin                   12.5 mg/ml (4 °C)                                 12.5 (μg/ml) (1:1000)*
		+	*Hygromycin                   50 mg/ml (4 °C)                                 125 (μg/ml) (:400)

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Latest revision as of 17:19, 9 September 2010

Concentrations for antibiotics

Antibiotic Stock E.coli B.subtilis

• Ampicillin (Amp) (in H2O) 50 mg/ml (4 °C) 100(μg/ml) (:500)
• Kanamycin (Km) 10 mg/ml (4 °C) 20 (μg/ml) (:500) 10 (μg/ml) (:100)
• Spectinomycin (Spec) (in H2O) 10 mg/ml (4 °C) 100(μg/ml) (:100) 100 (μg/ml) (:100)
• Erythromycin (Ery) E.coli stock 10 mg/ml (-18 °C) 150 (μg/ml) (:66.6)
• Erythromycin(Ery) B.subtilis stock 1 mg/ml (-18 °C) 2-5 (μg/ml)
• Tetracyclin (Tet) (50% EtOH) 3 mg/ml (-18 °C) 12 (μg/ml) (:250) 6(μg/ml) (:500)
• Chloramphenicol (Cm) (50% EtOH) 5 mg/ml (-18 °C) 10-15 (μg/ml) (:500/333) 5(μg/ml) (:1000)
• Lyncomycin 12.5 mg/ml (4 °C) 12.5 (μg/ml) (1:1000)*
• Hygromycin 50 mg/ml (4 °C) 125 (μg/ml) (:400)