Mgpure Viral RNA Purification Kit

Purpose: This procedure describes the extraction of viral RNA from virus infected specimen for real-time RT-PCR detection.

Acceptable Specimens

• Respiratory specimens including: nasopharyngeal or oropharyngeal aspirates or washes, nasopharyngeal or oropharyngeal swabs, broncheoalveolar lavage, tracheal aspirates, and sputum. o Swab specimens should be collected only on swabs with a synthetic tip with aluminum or plastic shafts. Swabs with calcium alginate or cotton tips with wooden shafts are not acceptable
• Serum, plasma, or other body fluid.

Viral RNA Extraction Protocol

1. Pipet 10 µL Proteinase K, 2 µL L Solution, 250 µL Buffer MYE and 500 µL isopropanol to each well of the 96 well sample plate. Note: Calculate number of samples and make master mix of proteinase K and Buffer MYE.
2. Add 250 µL sample to each well and mix well by pipetting. Incubate at room temperature for 10 min.
3. Add 10 µL MgPure beads to each well, mix well by pipetting and incubate at room temperature for 5 min.
4. Place the sample plate on the magnetic rack to magnetize the MgPure beads until the beads are completely cleared from the solution.
5. Remove the clear solution from the beads. Take the sample plate off the magnetic rack and resuspend the beads with 500 µL Buffer RB. Place the sample plate on the magnetic rack to magnetize the beads, remove the clear solution when the beads are completely cleared from the solution.
6. Take the sample plate from the magnetic rack and resuspend the beads with 500 µL RNA Wash Buffer. Place the sample plate on the magnetic rack to magnetize the beads, remove the clear solution when the beads are completely cleared from the solution.
7. Air-dry the sample for 5-10 minutes. Take the sample plate from the magnetic rack and resuspend the beads with 15-30 µL DEPC-treated H₂O. Place the sample plate on the magnetic rack to magnetize the beads, transfer the clear solution to a Elution Plate when the beads are completely cleared from the solution. Store RNA at -20℃ or put on ice for downstream applications.