Botrytis cinerea Detection Kits

PDF-116

Botrytis cinerea Detection Kits

Botrytis cinerea PCR Kit is designed for the detection of Botrytis cinerea specific DNA in a real-time PCR.

Botrytis cinerea PCR Kit is designed for the detection of B. cinerea specific DNA in a real-time PCR based on the use of technology. This kit is designed for research use only and not for use in diagnostic procedures. The detection of B. cinerea specific DNA is based on PCR providing a simple, reliable and rapid result for the detection of B. cinerea infection. Botrytis cinerea PCR Kit includes a PCR control to monitor for PCR inhibition, and to validate the quality of the sample and the detection result. The B. cinerea PCR Kit comprises Master Mix for the target and PCR control detection, Primer & Probe Mix, as well as a positive control and a negative control (nuclease-free water) to confirm the integrity of the kit reagents.

1. Before use, suitable amounts of all components should be completely thawed at room temperature, mixed by gentle vortexing or by pipetting, and centrifuged briefly.
2. Work quickly on ice.
3. The recommended minimum number of DNA samples tested per PCR run is 6.

Fungi

1. Follow universal precautions. All specimens should be considered as potentially infectious and handled accordingly.
2. Ensure that a suitable lab coat, disposable gloves, and protective goggles are worn when handling specimens and kit reagents.
3. Use sterile pipette tips with filters. Use proper pipetting techniques and maintain the same pipetting pattern throughout the procedure to ensure optimal and reproducible values.
4. Ensure that appropriate specimen collection, transport, storage, and processing techniques are followed for optimal performance of this test.

1. MDx 2X PCR Master Mix 350 µL
2. B. cinerea Primer & Probe Mix 70 µL
3. B. cinerea Positive Control 50 µL
4. Nuclease-Free Water (Negative control) 1.25 mL
5. Product Insert 1

1. Appropriate Real-Time PCR Instrument with FAM and HEX filter channel
2. DNA Purification Kit
3. Disposable powder-free gloves
4. Benchtop microcentrifuge
5. Micropipettors
6. Sterile pipette tips with filters
7. PCR tubes
8. Vortex mixer
9. PCR reaction preparation station (Optional)

Botrytis cinerea is a ubiquitous fungal pathogen, often present as a latent infection, which can in some cases later develops into damaging symptomatic infections in a wide range of plant species including economically important crops such as vegetables, ornamentals, bulbs and fruits. Consequently, an apparently healthy plant can deteriorate suddenly due to the development of latent or quiescent infection into visible disease. Sensitive and reliable methods to monitor the fungus in plant tissue are essential to enable the study of the factors affecting the progression from latent to aggressive disease, and to allow for early detection and diagnosis of the disease.

PCR

The specificity of Botrytis cinerea PCR Detection Kit is first and foremost ensured by the selection of the B. cinerea -specific primers, as well as the selection of stringent reaction conditions. The B. cinerea primers were checked for possible homologies to all microorganisms in GenBank published sequences by sequence comparison analysis.

All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots. All reagents can be stored for 1 year at -20°C without showing any reduction in performance.