Pseudomonas aeruginosa nucleic acid detection kit (fluorescent PCR method)|BNCC361453 |BNCC

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Pseudomonas aeruginosa nucleic acid detection kit (fluorescent PCR method)

Culture medium	Nutritional gravy medium (English name: NA/NB): beef paste 3.0g, peptone 10.0g,NaCl 5.0g, agar 20.0g (not included in liquid medium), distilled water 1.0L,pH 7.0. Sterilization at 121 ℃ for 15min. [Note] Adding 5 mgMnSO4 · H2O to culture Bacillus is beneficial to produce spores.
Subculture procedure	Experimental preparation: Before the experiment, the sample was cultured with reference to T/SDAQI 005-2021; the various components in the kit were dissolved on ice, slightly mixed and temporarily centrifuged, and then placed on ice for later use; Reagent preparation: experimental reaction system (25 μL),qPCR mixed reaction solution 18.5 μL, template 1 μL, sterile ddH2O5.5μL Reaction conditions: 95 ℃, 1min, 1cycle;95 ℃, 10s, 50 ℃, 30s, 40cycle; Fluorescence channel: FAM.
Growth conditions	Culture temperature 37 ℃; Culture time 18-24 hours; Aerobic gas environment;
Storage conditions	-20 ℃, cold storage
Safety level	2
Separation substrate	Blood culture
application	It is used for rapid detection of Pseudomonas aeruginosa in food and cosmetics.
Sharing mode	Public welfare sharing

Certificate of Pseudomonas aeruginosa Nucleic Acid Detection Kit

1. Product information

Sample name: Pseudomonas aeruginosa nucleic acid detection kit (fluorescence PCR method)

Sample number: 361453

Sample batch: 2200505

2. Product features

Description	Packaged in 1.5mL cryovials, containing reaction solution, positive quality control material, negative quality control material, ddH2O
Traceability	ATCC27853 Pseudomonas aeruginosa, ATCC23483 Pseudomonas putida
Product specifications	1 tube of reaction solution, 1 tube of positive quality control product, 1 tube of negative quality control product, 1 tube of ddH2O water
Field of application	Suitable for rapid detection of Pseudomonas aeruginosa in food and cosmetics
*Note: If you have any questions about this kit, please contact our center (BNCC) for help before use

3. Instructions

Sample processing: Refer to "SN/T 5228.9-2019 Rapid Screening Method of Pathogenic Microorganisms in Exported Foods for Pseudomonas aeruginosa" for enrichment culture; Experiment preparation: Dissolve various components in the kit on ice before use, slightly Mix well, centrifuge briefly, and place on ice for later use; Reagent preparation: experimental reaction system (25 μL), 18.5 μL of qPCR mixed reaction solution, 1 μL of template, 5.5 μL of sterile ddH2O; Reaction conditions: 95℃ for 1min 1cycle; ℃ 30s 40cycle; Fluorescence channel: FAM.

4. Notes

1. Sample processing, reagent preparation, and sample addition should be carried out in different areas to avoid cross-contamination;

2. To ensure the sensitivity of the detection, it is recommended that the qPCR mixed reaction solution be frozen and thawed no more than 3 times;

3. Judgment of results: if the Ct value of the test sample is greater than or equal to 40 and there is no typical amplification curve, it is judged as negative; if the Ct value of the test sample is less than or equal to 40 and there is a typical amplification curve, it is judged as positive; if the test sample is tested Ct values >35 and <40 suggest that the sample be redone.

5. Preservation and transportation

long-term storage:-20 ℃, valid for 1 year; Transportation conditions: dry ice transportation.

Henan Engineering Technology Research Center of Industrial Microbial Strain

website: www.bncc.org.cn tel: 400-6699-833

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