Magni:Protocols/staining

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(New page: {{Magni}} <div style="padding: 10px; width: 720px; border: 5px solid #000000;"> = Blue-white screening = This screening is to select colonies expressing a functional Beta-galactosidase pr...)
Current revision (09:46, 31 October 2012) (view source)
 
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*Pre-warm an LB agar plate at 37degC.
*Pre-warm an LB agar plate at 37degC.
-
*For each plate, mix 20 ul of X-Gal (40 mg/ml dissolved in DMF), 20 ul of IPTG (200 mM, ReadyMade stock) and 60 ul of sterile LB.
+
*For each plate, mix 20 ul of X-Gal (40 mg/ml dissolved in DMF), 20 ul of IPTG (200 mM, ReadyMade stock) and 160 ul of sterile LB.
*Spread evenly on the plate near the flame.
*Spread evenly on the plate near the flame.
*Incubate it at 37degC until dry and use.
*Incubate it at 37degC until dry and use.

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Blue-white screening

This screening is to select colonies expressing a functional Beta-galactosidase protein (lacZ gene). They appear as blue colonies on agar plates.

  • Pre-warm an LB agar plate at 37degC.
  • For each plate, mix 20 ul of X-Gal (40 mg/ml dissolved in DMF), 20 ul of IPTG (200 mM, ReadyMade stock) and 160 ul of sterile LB.
  • Spread evenly on the plate near the flame.
  • Incubate it at 37degC until dry and use.

SAFETY NOTE: DMF is toxic, always use gloves and a face mask.

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