Introduction

This kit is suitable for the detection of Escherichia coli O157:H7 in food. Based on Real Time PCR technology, using primers, fluorescent probes and other reagents required for the specific gene of Escherichia coli O157:H7, the amplification reaction can be performed by adding the sample to be tested.

Principle

During the amplification process, the fluorescent probe binds to the target gene fragment, which can be decomposed by Taq enzyme and produce a fluorescent signal. At this time, the fluorescent quantitative PCR instrument can recognize the fluorescent signal and draw the corresponding real-time amplification curve according to its intensity changes, thereby determining whether Escherichia coli O157:H7 is detected.

1. Sample Pretreatment

• Sample enrichment: Take 25 g (mL) of sample, add to 225 mL modified EC broth, homogenize, and culture at 36±1°C for 18-24 h.
• Centrifugation: Pipette 1 mL enrichment solution, centrifuge at 6000 r/min for 5 min, discard supernatant. Wash with 50 μL sterile ultrapure water.
• Lysis: Add 30 μL lysis solution, suspend bacteria, and heat at 99°C for 10 min.
• Final Preparation: Centrifuge at 12000 r/min for 15 min. The supernatant is the crude DNA template.

2. Sample Addition and Reaction

• Prepare PCR reaction tubes (Negative control + Samples + Positive control).
• Add 20 μL of premix to each tube.
• Add 5 μL of template DNA (or control) to the tubes.
• Perform PCR amplification in the FAM detection channel.

• Offering Samples for Testing.
• R & D team providing technical support.
• Provide packaging and labeling OEM customization.