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Summary: 1
Bacterial Colony Hybridization
But : Identifier parmi des centaines de colonies, les colonies positives, c'est-à-
dire les bactéries qui ont incorporé l'insert désiré.
Material and reagents :
- Filter #1 (trim to fit in petri dishes as necessary). The filters do not need to be
sterile.
- Coomassie brilliant blue R-250, powder.
- Sonde spécifique.
Reagents to prepare :
50 mM NaOH, 0.5% SDS (solution A) 200 ml
NaOH 10 N 1 ml
SDS 20% 5 ml
DdH2O 194 ml
Hybridization mix 10 ml
20X SSC ou SSPE 2.5 ml (5X SSC)
20% SDS 250 µl (0.5% SDS)
Formamide 5 ml (50% Form.)
1 M Tris, pH 7.5 0.5 ml (50 mM Tris)
1 M EDTA 50 µl (5 mM EDTA)
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