16S/18S FISH Probe Design & Synthesis
Description
The 16S and 18S rRNA sequences are the preferred targets for members of the bacterial/archaeal and eukaryotic domains because they are universally present and highly abundant, consisting of highly conserved regions and variable regions. The FISH technique utilizes these gene sequences to design specific probes that can accurately locate and identify target microorganisms. This provides a powerful tool for analyzing microbial community structures, studying ecological functions, and diagnosing diseases. Probe design is indeed a critical step that can affect the performance of FISH. After years of development, we have launched a species-specific 16S/18S FISH probe design, synthesis, and detection service, offering researchers an unparalleled 16S/18S FISH probe experimental experience.
Selection of Fluorescent Labels for FISH Probes
The table below lists the wavelength ranges of the filter sets for the four commonly used fluorescent channels in fluorescence microscopy: DAPI, FITC, Texas Red, and CY5. It also includes the types of fluorescent groups that are most compatible with these channels, along with their primary applications (such as antibody labeling, protein expression, or FISH probe labeling):
| Filter Set | Excitation Filter (nm) | Emission Filter (nm) | Best Matching Fluorophore Types | Compatible Fluorophore Types | Main Applications |
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Please note that these values and the recommended fluorophores are based on common fluorescence microscope filter settings and the characteristics of fluorescent dyes. In practical applications, it is advisable to refer to the microscope’s user manual, the specifications from filter manufacturers, and information from fluorescent dye suppliers to ensure optimal fluorescence signal and imaging results.
Recommended FISH Probe Fluorescent Labeling Strategies (Please confirm based on actual conditions):
Single Bacterium 16S FISH Labeling:
- Target Bacterium Labeling: Preferably label with Texas Red, compatible with CY3
- Negative Control: Recommended to label with FITC (*Microbial FISH services can provide this for free)
- Positive Control (Universal Bacterial Probe): CY5
- DAPI Channel: Stains DNA
Dual-Color FISH Probe Labeling (Simultaneously detecting two bacteria, or using two different fluorescent labels to detect the same bacterium):
- Positive Control (Universal Bacterial Probe): FITC
- Target Bacterium 1: Recommended to label with Texas Red, compatible with CY3
- Target Bacterium 2: Recommended to label with CY5
- DAPI Channel: Stains DNA
In this case, if a negative control is needed, it should be tested on a separate slide.
Product Features
- Each probe is rigorously designed and evaluated to ensure high specificity and high coverage.
- Each probe comes with a detailed evaluation report, including specificity, coverage, and application recommendations.
- For single FISH probes with poor specificity, dual probes and multiple probes are designed to improve specificity.
Benefits of Using Dual Probes in FISH 16S rRNA Detection:
- Increased Specificity: Since the 16S rRNA gene has different conserved and variable regions in different bacteria, using dual probes can simultaneously target these regions, thereby enhancing the specific identification of the target species.
- Improved Sensitivity and Accuracy: Dual probes provide double confirmation, reducing false positives caused by potential non-specific binding of a single probe.
- Wide Applicability: Dual-probe FISH technology can be applied to the detection of various microorganisms, including bacteria, archaea, and fungi, making it a versatile tool for microbial detection.
- Aids in Microbial Community Analysis: In microbial community structure analysis, dual probes can more accurately identify and distinguish different members of the community, providing important information for microbial ecology research.
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