QIAmp DSP Virus Kit DNA/RNA Extraction Procedure

The QIAamp DSP Virus procedure combines the selective binding properties of a silica-based membrane with minimal elution volumes of 20 ul or 60 ul.


Principle:
The QIAamp DSP Virus procedure comprises 4 steps:

• Lysing the virus particles in the sample 
• Binding the viral nucleic acids in the lysate to the membrane of a QIAamp Minelutae Column 
• Washing the membrane 
• Eluting the viral nucleic acids from the membrane 

DNA/RNA Extraction Procedure:

• Pipet 75 ul QIAGEN Protease (QP) into a lysis tube. 
• Add 500 ul of plasma or serum to the tube. 
• Add 500 ul lysis buffer/RNA CARRIER and mix by pulse-vortexing for 15 sec 
• Incubate at 56°C for 15mins. 
• Centrifuge for ≤5 seconds at full speed to remove drops from the inside of lid. 
• Change gloves and add 600 ul ethanol, mix by pulse-vortexing. Incubate for 5 minutes at room temperature then centrifuge at full speed. 
• Change gloves & transfer the lysate into the QIAmp mini spin column 
• Centrifuge for 1 min at 12,000 rpm. 
• Discard the filtrate & replace with new clean 2 ml tube. 
• Apply 600 ul Wash buffer 1 to the MinElute column and spin for 1 min at 12,000 rpm for 3 min.
• Discard filtrate and replace with a new 2 ml tube. 
• Add 750 ul of Wash Buffer 2 and spin for 5 minutes at 14,000 rpm. 
• Discard filtrate and replace with new tube, add 750 ul ethanol and spin for 1 min at 14,000 rpm. Discard tube and replace with a new 2 ml tube. Do an extra spin. 
• Incubate spin column at 56ºC for 3 mins with the lid open to evaporate remaining liquid. 
• Place the spin column in a clean tube & discard the filtrate. Add 60 ul elution buffer and incubate for 5 mins at room temperature. 
• Spin the tube for 1 min at 14,000 rpm to elute the viral nucleic acids.
• Discard MinElute Column and sample is now ready for PCR.