RNA extractions for both methods were carried out using the MagMAX™ Pathogen RNA/DNA Kit (Applied Biosystems®) on a MagMAX™ Express 96 Extraction Robot (Applied Biosystems®) with a sample input of 200 μL, and elution volume of 90 μL. One μL per reaction of VetMAX™ Xeno™ Internal Positive Control RNA (10,000 copies/μL) (Applied Biosystems®) was also added to the lysis buffer prior to extraction.
Vetmax xeno internal positive control rna
Manufactured by Thermo Fisher Scientific
The VetMAX™ Xeno™ Internal Positive Control RNA is a laboratory product designed to be used as an internal positive control in molecular diagnostic assays. It is intended to verify the performance of the assay and the integrity of the sample during the testing process.
Automatically generated - may contain errors
2 protocols using vetmax xeno internal positive control rna
1
Automated RNA Extraction for Pathogen Testing
2
Foot-and-Mouth Disease Virus Detection in Milk
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Diagnostic screening methods for the detection of FMDV genome in milk samples are defined as Method A and Method B for the purpose of this study, and are described in Table 2 . In initial studies, the performance of four RNA extraction and rRT-PCR combinations was assessed. However, some reagents/extraction robots are no longer commercially available or used in diagnostic laboratories, therefore only two methods (A and B) were carried forward for further evaluation in this study.Table 2
Comparison of the two high-throughput foot-and-mouth disease virus detection methods.
| A | B | |
|---|---|---|
| Extraction kit | MagMAX™ Pathogen RNA/DNA Kit (Applied Biosystems®) | MagMAX™ Pathogen RNA/DNA Kit (Applied Biosystems®) |
| Internal Control | VetMAX™ Xeno™ Internal Positive Control RNA (Applied Biosystems®) | VetMAX™ Xeno™ Internal Positive Control RNA (Applied Biosystems®) |
| Sample input | 200 μL | 200 μL |
| rRT-PCR kit | ‘TaqMan® Fast’ Virus 1-Step Master Mix (Applied Biosystems®) | ‘Superscript’ III Platinum® One-Step qRT-PCR Kit (InvitrogenTM) |
| Internal control assay | VetMAX™ Xeno™ Internal Positive Control LIZ™ Assay (Applied Biosystems®) | VetMAX™ Xeno™ Internal Positive Control LIZ™ Assay (Applied Biosystems®) |
| Primers and Probes | Targeting 3D polymerase (Callahan et al., 2002 (link)) | Targeting 3D polymerase (Callahan et al., 2002 (link)) |
| RNA template input | 2.5 μL | 5 μL |
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