Magni:Protocols/staining: Difference between revisions
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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...) |
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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 | *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. | ||
Latest revision as of 06:46, 31 October 2012
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.
