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Applied and Environmental Microbiology, April 2002, p. 2093, Vol. 68, No. 4
0099-2240/02/$04.00+0 DOI:
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
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Cardiff School of Biosciences, Cardiff University, Cardiff CF10 3TL, United Kingdom
Volume 68, no. 1, p. 201-210, 2002. Page 203, column 1 for 2 h in 5x SSC-2% blocking solution (Boehringer Mannheim)-0.1% N-lauryl sarcosine-0.02% SDS. Hybridization was carried out for 12 to 18 h in hybridization buffer containing 0.9 M NaCl-0.1% N-lauryl sarcosine-4% blocking reagent-0.01% SDS-formamide (20, 30, 40, 50, and 60%)-20 ng of probe ml-1. Stringency washes were carried out twice for 15 min in 0.01% SDS-0.02 M Tris-HCl (pH 7.4)-NaCl (0.19, 0.074, 0.037, 0.019, and 0.013 M). Optimal hybridization conditions were chosen when there was no detection for nontarget organisms and substantial detection for target CFBs. Optimal hybridization conditions for probes CFB560, CFB562, and CFB376 were found at 40.
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