rRNA targeted fluorescent in situ hybridization analysis of bacterial community structure in river water

Article Abstract:

A ribosomal RNA fluorescent in situ hybridization approach using 2-hydroxy-3-naphthoic acid 2'-phenylanilide phosphate and Fast Red TR was developed to study planktonic bacterial cells in river water. The bacterial community in two sites with different organic carbon pollution level was investigated using oligonucleotide probes specific for the domain bacteria and five bacterial groups. Results showed that while the probes were able to identify 50% of the total cells at the eutrophic sites, only 26-38% of the cells were identified at the oligotrophic site.

Nucleic acid probes, Ribosomal RNA, Water pollution, Rivers, Plankton populations

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Detection of specific bacterial cells with 2-hydroxy-3-naphthoic acid-2'-phenylanilide phosphate and Fast Red TR in situ hybridization

Article Abstract:

An in situ hybridization technique using 2-hydroxy-3-naphthoic acid-2'-phenylanilide phosphate (HNPP) and Fast Red TR is used to effectively identify single bacterial cells. The reaction of the dephosphorylated form of HNPP with Fast Red TR produces fluorescent signals. The signals are strong and last for almost two hours, permitting exact observation and counting. The method provides the effective detection of low-rRNA-content or small bacterial cells in nature.

Observations, In situ hybridization, Fluorescence

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Recognition of individual genes in diverse microorganisms by cycling primed in situ amplification

Article Abstract:

The cycling primed in situ amplification-fluorescent in situ hybridization (CPRINS-FISH) is developed to recognize individual genes in a single bacterial cell. The results demonstrate that CPRINS-FISH provides a high sensitivity for microscopic detection of bacteria carrying a specific gene in natural aquatic samples.

Science & research, Genetic aspects, Microorganisms, Hybridization

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