Introduction
Purify viral RNA purification from viral transport medium (VTM) using a fully automated protocol with the Maxwell® HT Viral TNA Kit, Custom on the CyBio FeliX liquid handler.
Kit: Maxwell® HT Viral TNA Kit, Custom (Cat.# AX2340)
Analyses: RT-qPCR
Sample Type(s): Samples collected in viral transport medium, e.g. nasopharyngeal swabs
Input: 200 µL
Materials and Methods
Materials and Methods
Instruments and equipment
- CyBio FeliX Basic Unit with Enclosure (Cat.# OL5015-24-100)
- CyBio FeliX Extraction Set (Cat.# OL5015-25-120)
- Protective Plate (Cat.# OL3317-25-126)
- Heat block
- Microcentrifuge
Consumables and reagents
- Maxwell® HT Viral TNA Kit, Custom (Cat.# AX2340)
- CyBio TipRack 96/1000 µL, PCR-certified, pre-sterilized, filter (Cat.# OL3811-25-939-F)
- Deep Well Plate, 96 square well, 2 mL (Cat.# 845-FX-8500025)
- 100% Isopropanol (molecular biology grade)
- 100% and 80% Ethanol (molecular biology grade)
Method
1. Add Isopropanol to 4/40 Wash Solution, as indicated on the bottle.
2. Add 100% Ethanol and Isopropanol to Alcohol Wash, as indicated on the bottle.
3. Prepare 80% Ethanol.
4. Prepare a lysis/binding mastermix for all samples plus 10% overage in an appropriately sized container. For each sample, add 150 µL of Lysis Solution, 15 µL of Proteinase K, 300 µL of 100% Isopropanol, and 15 µL of
MagneSil RED. Invert the mixture until the contents are homogeneous (do
not vortex).
5. For each sample to be processed, add 480 µL of the lysis/binding mastermix from step 4 to wells of the 96 well processing plate.
6. Add 200 µL of sample to the 96 well plate processing plate containing the lysis/binding mastermix.
7. The method uses the following reagents and volumes per well (Table 1).
8. Start the run on the CyBio Felix Liquid Handler – Maxwell HT Viral TNA_Version1.bms.
9. Follow the instructions on the instrument for reagent setup. The instrument deck configuration is shown in Figure 1. The user will be prompted during the method start to load the sample plate containing Samples & Lysis/Bind reagents on the BioShake (position 1 on deck layout). The automated method can be configured with a variable elution volume. This Application Note was performed with 60 µL and 110 µL as elution volumes.
Results
