Agilent Technologies Porcine Detection Kit User Manual
Summary
A comprehensive guide for testing food products and samples for the presence of porcine DNA using quantitative PCR (qPCR) analysis. This manual provides detailed protocols for genomic DNA extraction, QPCR amplification, result interpretation, and troubleshooting. Ideal for laboratory researchers and quality control professionals in the food safety industry, it ensures proper sample handling and decontamination procedures are followed.
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Porcine Detection Kit 5500-0094
Protocol Version A, April 2011 Research Use Only. Not for use in Diagnostic Procedures.
Agilent Technologies
Page Summary Contents For Agilent Technologies Porcine Detection Kit User Manual
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Porcine Detection Kit 5500-0094 Protocol Version A, April 2011 Research Use Only. Not for use in Diagnostic Procedures. Agilent Technologies
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Notices © Agilent Technologies, Inc. 2011 Warranty Safety Notices No part of this manual may be reproduced in The material contained in this any form or by any means (including elec- document is provi...
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In this Guide... This document describes how to use the Porcine Detection Kit to extract genomic DNA from a food sample or other product and test it for the presence of porcine DNA using QPCR analysis...
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Porcine Detection Protocol
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Contents Before You Begin 7 Kit Contents 8 Materials Provided with the Agilent Porcine Detection Kit 8 Storage Conditions 8 Required Equipment, Supplies and Reagents 9 Overview of the Agilent Porcine ...
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Contents If the porcine-specific target is amplified in the NTC reactions 26 If the porcine-specific target fails to amplify in the positive control reactions 26 If the Alien target fails to amplify i...
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Porcine Detection Kit Protocol Before You Begin Kit Contents 8 Materials Provided with the Agilent Porcine Detection Kit 8 Storage Conditions 8 Required Equipment, Supplies and Reagents 9 Overview of ...
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Before You Begin Kit Contents Kit Contents Materials Provided with the Agilent Porcine Detection Kit DNA Isolation Kit, Part Number 5500-0051 (Boxes 5972-3629 and 5972-3630), 50 preparations Nucleic A...
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Before You Begin Required Equipment, Supplies and Reagents Required Equipment, Supplies and Reagents Additional Equipment, Supplies and Reagents Required Equipment for food and beverage homogenization...
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Before You Begin Overview of the Agilent Porcine Detection Kit and Protocol Overview of the Agilent Porcine Detection Kit and Protocol The Agilent Porcine Detection Kit contains reagents and materials...
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Before You Begin The Agilent Porcine Detection QPCR Kit provided in the porcine detection assay mix. The master mix has been optimized for maximum performance on Agilent’s Mx3000P and Mx3005P QPCR sys...
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Before You Begin Preprotocol Considerations Preprotocol Considerations Avoiding Cross-Contamination Take food samples that are to be tested straight to the laboratory to be weighed in labeled containe...
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Before You Begin Avoiding Cross-Contamination that is in an entirely separate room from the other two areas. The extraction procedure is the step that is most likely to generate aerosols that can lead...
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Before You Begin Avoiding Cross-Contamination Porcine Detection Protocol
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Porcine Detection Kit Protocol Procedures Preparations 16 To prepare the reagents 16 To prepare the test samples 16 DNA Extraction and Isolation 18 To extract DNA 18 To isolate DNA 18 QPCR Amplificati...
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Procedures Preparations Preparations To prepare the reagents High Salt Wash Buffer Prepare 1× High Salt Wash Buffer by adding 16 ml of 100% ethanol to the bottle of High- Salt Wash Buffer. After addin...
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Procedures To prepare the test samples Suggested Homogenization Methods If a large volume is to be homogenized – e.g. a can of soup or multiple slices of meat – place the sample in a beaker (add a suf...
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Procedures DNA Extraction and Isolation DNA Extraction and Isolation To extract DNA Pre- warm the Proteinase K Digestion Buffer to 65°C for 5 minutes in an incubator or water bath; invert to mix. Prep...
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Procedures To isolate DNA Remove and retain the spin cups and discard the filtrates. For each sample, replace the spin cup in the receptacle tube, then add 500 μl of 1× High Salt Wash Buffer and cap t...
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Procedures QPCR Amplification QPCR Amplification Please read the following notes carefully before beginning the QPCR set up: Once the QPCR reagents are thawed (the 2× master mix, the porcine detection...
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Procedures To set up the QPCR reactions Example: If you will be running 64 reactions, prepare enough reagent mixture for at least 65 reactions (and preferably more). Table 1 PCR Reagent Mixture Compon...
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Procedures To run the QPCR protocol You do not need to mix the reactions after adding DNA. Mixing can create bubbles, which interfere with fluorescence detection. To run the QPCR protocol If possible,...
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Procedures To interpret the QPCR results If the porcine- specific target is amplified in the NTC reactions, then your reactions have been contaminated. If the Alien target fails to amplify in any of t...
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Procedures To interpret the QPCR results The exact Ct of the porcine- specific target is dependent on the quantity of porcine DNA present in the sample extract (higher levels of porcine DNA result in ...
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Porcine Detection Kit Protocol Troubleshooting Guide Troubleshooting 26 This chapter offers recommendations for dealing with potential issues that may arise when using the Agilent Porcine Detection ki...
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Troubleshooting Guide Troubleshooting Troubleshooting If the porcine-specific target is amplified in the NTC reactions The reactions are contaminated. Repeat the experiment using a fresh set of QPCR r...
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Troubleshooting Guide If the Alien target is amplified poorly in the test reactions compared to the control reactions (i.e. more than 2 cycles later) Something in the sample extracts is inhibiting PCR...
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Troubleshooting Guide If the Alien target is amplified poorly in the test reactions compared to the control reactions (i.e. more than 2 cycles later) Porcine Detection Protocol
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Porcine Detection Kit Protocol Appendix To decontaminate an immersion blender 30 This chapter contains instructions on decontaminating the immersion blender used for sample homogenization to prevent c...
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Appendix To decontaminate an immersion blender To decontaminate an immersion blender If using an immersion blender for sample homogenization, decontaminate the blender after each use. The instructions...
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www.agilent.com In This Book This document describes how to use the Porcine Detection Kit to extract genomic DNA from a food sample or other product and test it for the presence of porcine DNA using Q...
Manual Details
| Brand | Agilent Technologies |
|---|---|
| Pages | 32 |
| File Size | 491.93 KB |
| Published | May 13, 2026 |
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