The use of fluorescein for labeling genomic probes in the checkerboard DNA-DNA hybridization method

被引：10

作者：

do Nascimento, Cassio ^{[1
]}

Santos Barbosa, Rodrigo Edson ^{[1
]}

Mardegan Issa, Joao Paulo ^{[1
]}

Watanabe, Evandro ^{[2
]}

Ito, Izabel Yoko ^{[2
]}

Monesi, Nadia ^{[2
]}

de Albuquerque Junior, Rubens Ferreira ^{[1
]}

机构：

[1] Univ Sao Paulo, Dept Mat Dentarios & Protese, Fac Odontol Ribeirao Preto, BR-14040904 Sao Paulo, Brazil

[2] Univ Sao Paulo, Dept Anal Clin Toxicol & Bromatol, Fac Ciencias Farmaceut Ribeirao Preto, BR-14040903 Sao Paulo, Brazil

来源：

MICROBIOLOGICAL RESEARCH | 2008年 / 163卷 / 04期

基金：

巴西圣保罗研究基金会;

关键词：

fluorescein; chekerboard technique; genomic probes;

D O I：

10.1016/j.micres.2006.11.020

中图分类号：

Q93 [微生物学];

学科分类号：

071005 [微生物学]; 100705 [微生物与生化药学];

摘要：

Molecular methods that permit the simultaneous detection and quantification of a large number of microbial species are currently employed in the evaluation of complex ecosystems. The checkerboard DNA-DNA hybridization technique enables the simultaneous identification of distinct bacterial. species in a large number of dental samples. The original technique employed digoxigenin-labeled whole genomic DNA probes which were detected by chemiluminescence. In this study, we present an alternative protocol for labeling and detecting whole genomic DNA probes in the Checkerboard DNA-DNA hybridization method. Whole genomic DNA was extracted from five bacterial species and labeled with fluorescein. The fluorescein labeled whole genomic DNA probes were hybridized against whole genomic DNA or subgingival plaque samples in a checkerboard hybridization format, followed by chemiluminescent detection. Our results reveal that fluorescein is a viable and adequate alternative labeling reagent to be employed in the checkerboard DNA-DNA hybridization technique. (c) 2007 Elsevier GmbH. All rights reserved.

引用

页码：403 / 407

页数：5

共 23 条

[1]

Subgingival plaque microbiota in Saudi Arabians after use of miswak chewing stick and toothbrush [J].