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        ß - GAL Filter Assay

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        922

        ß - GAL Filter Assay

         

         

        • streak yeasts to single colony
        • replica plate onto NC-filter (leave 10 sec.)
        • use tweezers to lift filter and slowly submerge in liquid Nitrogen for 1 min
        • place on back side of a petri dish to thaw (caution filter is very brittle !)
        • take new petri dish lid, add 2 ml Z-buffer
        • add 35 µl X-GAL solution directly into the Z-buffer, mix well
        • set Whatman filter disk (9 cm) into lid to absorb buffer
        • lay thawed filter on top , avoid bubbles
        • place in 30 °C incubator
        • incubate 10 - 60 min

         


        Buffers:

         Z-buffer:
        60 mM Na
        2 HPO 4 (anhyd.); 60 mM NaH 2 PO 4 ; 10 mM KCl; 1 mM MgSO 4
        [for 500 ml: 4.26 g Na 2 HPO 4 (anhyd.); 2.40 g NaH 2 PO 4 ; 373 mg KCl; 123 mg MgSO 4 ; adjust to pH 7.0]

         X-GAL solution:
        40 mg/ml in DMF (dimethylformamide)

         

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