96-well Viral DNA/RNA Kit

Details for Product No. ABIN5954823,
Application
DNA Extraction (DEx), RNA Extraction (REx)
Options
Purpose An easy and reliable method for isolating total viral DNA/RNA from whole blood in a 96-well format.
Sample Type Blood
Characteristics Features and Benefits: An easy and reliable method for isolating total viral DNA/RNA from whole blood in a 96-well format.
Components
  • 96-Well DNA/RNA Plate
  • 96-Well Deep Plates
  • Buffer HLY
  • Wash Buffer*
  • Protease K
  • Carrier Nucleic acid
  • Sealing Film
  • DEPC Water
*Add 200 mL 100% ethanol to RNA Wash Buffer before use
Material not included
  • Centrifuge capable 4,000g with adaptor for 96-well plates at 4 -22 °C.
  • Vortex
  • Multichannel pipet.
  • RNase-free filter tips
  • Reagent reservoirs for multichannel pipet
  • RNase-free 1.2 mL 96-well plates
  • 2 mL 96-well plates
  • 100 % ethanol
Application Notes The kit is also suitable for isolation of total RNA from cultured cells, tissues, and bacteria. This procedure has been tested for isolating nucleic acids from Hepatitis A, Hepatitis C and HIV. The isolated DNA/RNA can be used for PCR, RTPCR and other downstream applications
Comment

Purified DNA is suitable for PCR, restriction digestion, and hybridization techniques

Reagent Preparation
  1. Add 200 mL 100 % ethanol to RNA Wash Buffer before use.
  2. Add all Carrier Nucleic acid to Buffer HLY. Transfer Carrier Nucleic acid to the bottle of HLY Lysis Buffer. Store at 4 °C. Warm up Buffer HLY/carrier before use if precipitation forms at 4 °C.
  3. Prepare Wash Buffer and HLY Lysis Buffer/carrier Nucleic acid solution as instructed
Assay Procedure
  1. Add 150 μL plasma, acellular body fluid, cell culture supernatant to each well of a 1.2 mL 96-well plate (not provided).
  2. Add 10 μL Protease K to each well.
  3. Add 500 μL HLY Lysis Buffer/Carrier Nucleic acid solution to each well. Seal the plate with a Sealing Film.
  4. Keeping the 96-well plate flat on the bench, shake vigorously back and forth for 30 seconds. Rotate the plate 90° and shake the plate for another 30 seconds.
  5. Let sit at room temperature for 10 minutes.
  6. Briefly centrifuge at 500g to collect any liquid droplets from the film.
  7. Remove and discard the Sealing Film.
  8. Add 350 μL 100 % ethanol to each well. Seal the plate with a Sealing Film.
  9. Vortex the plate for 30 seconds. Briefly centrifuge at 500g to collect any liquid droplets from the film. Remove and discard the Sealing Film.
  10. Place a 96 Well Plate onto a 96-well Deep Plate (provided).
  11. Transfer 500 μL sample from Step 8 (including any precipitate that may have formed) to each well of the 96 Well Plate.
  12. Seal the 96 Well Plate with a Sealing Film.
  13. Centrifuge at 4,000g for 5 minutes at room temperature.
  14. Discard the filtrate from the 96-Well Deep Plate.
  15. Remove and reuse the Sealing Film in the following step.
  16. Repeat Steps 11-15 until the entire sample has been transferred to the 96 Well Plate.
  17. Remove the Sealing Film.
  18. Add 750 μL Wash Buffer to each well. Seal the plate with the Sealing Film. Place the 96 Well Plate onto the 96-well Deep Well Plate.
  19. Centrifuge at 4,000g for 10 minutes at 4 °C. Discard the flow through and centrifuge the plate at 4000xg for 10 min at 4 °C.
  20. Note: It is important to dry the 96 Plate matrix before elution. Residual ethanol may interfere with downstream applications.
  21. Remove and discard the Sealing Film. Transfer the 96 Well Plate to an Elution Plate.
  22. Add 100-150 μL DEPC-treated water to each well. Seal the plate the new Sealing Film. Make sure to add water directly onto RNA matrix.
  23. Let sit for 1 minute at room temperature.
  24. Centrifuge at 4,000g for 5 minutes at 4 °C. The RNA/DNA is in the flow through.
  25. Store RNA/DNA at -80 °C.
Restrictions For Research Use only
Storage RT,4 °C,-20 °C
Storage Comment

Storage: RT

storageComment: All components of the 96-Well Viral DNA/RNA Kit should be stored at 22°C -25°C except the Buffer HLY, Carrier, Protease K. Buffer HLY, Carrier and Protease K should be stored at 2-8°C. DO NOT FREEZE!

Expiry Date 12 months