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and inexpensively produced. During oligonucleotide synthesis a variety of marker or linker sequences can be introduced to the 5' end of the oligonucleotide. The principal conjugate fluorochromes are derivatives of fluorescein or rhodamine, although labels such as digoxigenin and biotin have also been used (Fuhrman et al. 1994, Amann et al. 1995). The design of rRNA-targeted group- or species-specific probes should be based on a good rRNA sequence database and be performed in a computer-assisted way using appropriate software. The organisms encompassed by a probe varies according to the region of the 16S rRNA selected as the hybridization target. Species-specific probes complement the most variable regions, while more general probes target more conserved regions of the molecule. The principal steps involved in the design of probes are: the alignment of rRNA gene sequences; the identification of sequence idiosyncrasies; the synthesis and labelling of complementary nucleic acid probes; and the experimental evaluation and optimization of the probe specificities and assay sensitivities using cultured reference strains (Manz et al. 1992, Devereux et al. 1992).
DOI link for and inexpensively produced. During oligonucleotide synthesis a variety of marker or linker sequences can be introduced to the 5' end of the oligonucleotide. The principal conjugate fluorochromes are derivatives of fluorescein or rhodamine, although labels such as digoxigenin and biotin have also been used (Fuhrman et al. 1994, Amann et al. 1995). The design of rRNA-targeted group- or species-specific probes should be based on a good rRNA sequence database and be performed in a computer-assisted way using appropriate software. The organisms encompassed by a probe varies according to the region of the 16S rRNA selected as the hybridization target. Species-specific probes complement the most variable regions, while more general probes target more conserved regions of the molecule. The principal steps involved in the design of probes are: the alignment of rRNA gene sequences; the identification of sequence idiosyncrasies; the synthesis and labelling of complementary nucleic acid probes; and the experimental evaluation and optimization of the probe specificities and assay sensitivities using cultured reference strains (Manz et al. 1992, Devereux et al. 1992).
and inexpensively produced. During oligonucleotide synthesis a variety of marker or linker sequences can be introduced to the 5' end of the oligonucleotide. The principal conjugate fluorochromes are derivatives of fluorescein or rhodamine, although labels such as digoxigenin and biotin have also been used (Fuhrman et al. 1994, Amann et al. 1995). The design of rRNA-targeted group- or species-specific probes should be based on a good rRNA sequence database and be performed in a computer-assisted way using appropriate software. The organisms encompassed by a probe varies according to the region of the 16S rRNA selected as the hybridization target. Species-specific probes complement the most variable regions, while more general probes target more conserved regions of the molecule. The principal steps involved in the design of probes are: the alignment of rRNA gene sequences; the identification of sequence idiosyncrasies; the synthesis and labelling of complementary nucleic acid probes; and the experimental evaluation and optimization of the probe specificities and assay sensitivities using cultured reference strains (Manz et al. 1992, Devereux et al. 1992).
ABSTRACT
Quantitative Dot-blot Hybridization