DNA Transfer - Plaque/Colony Lift
1. Wet circular Nitrocellulose filter in TE and place on chilled agar plate. Orient the filter on the plate by puncturing the filter asymmetrically with a needle.
2. After 4 min carefully lift the filter from the plate and transfer it to Whatman 3MM paper saturated with:
0.5 M NaOH 10 mL 5 N NaOH
1.0 M NaCl 20 mL 5 M NaCl
70 mL ddH20
3. After 4 min, transfer the filter to Whatman 3MM paper saturated with 1 M Tris, pH 7.5.
4. After 4 min, transfer filter to Whatman 3MM paper saturated with:
0.5 M Tris, pH 7.5 50 mL 1 M Tris, pH 7.5
1.5 M NaCl 30 mL 5 M NaCl
20 mL ddH2O
5. Transfer to a piece of dry Whatman 3MM paper and air dry.
6. Bake at 80° for 90 min.
Last Update: Thursday, 19-Jun-2014 11:50:32 PDT
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