Viral RNA+DNA Preparation Kit - Izolace DNA a RNA

Detailní popis

For in vitro use only!

Shipping: shipped at ambient temperature

Storage Conditions: store at ambient temperature

Shelf Life: 12 months

Description:
Viral RNA+DNA Preparation Kit is designed for rapid and effective isolation of RNA and DNA from a variety of pathogen organisms such as virus or bacteria. Samples can be fresh or frozen plasma/blood (treated with anticoagulants excepted heparin), serum, other cell-free body fluids or pathogen-infected tissue. The Kit is specifically designed to isolate high-quality nucleic acids using low elution volumes and allowing sensitive downstream analysis including quantitative PCR and RT-PCR. The purified RNA/DNA is free of proteins and nucleases. Viral RNA+DNA Preparation Kit uses lysis buffer including chaotropic salts to inactivate RNases/DNases and advanced silica-gel membrane technology for fast purification of intact RNA/DNA. The preparation procedure is optimized to give reproducible results within 30 min.

Content:

Additional Materials Required:
96-99% Ethanol
1.5 ml Tubes

Preparation procedure:
1a Preparation from plasma, serum, urine, cell-culture supernantant, cell-free fluid or virus infected tissue

• Transfer 150 μl plasma, serum, urine, cell-culture supernatant, cell-free fluid or virus infected tissue into a 1.5 ml microtube.
• Note: Adjust lower sample volumes with PBS Buffer to 150 μl. Samples of larger volumes (up to 300 μl) can easily be scaled up but may require larger tubes for the lysis procedure.
• Add 300 μl (2 amounts of sample volume) of Lysis Buffer.
• Vortex for 15 sec.
• Incubate at room temperature (20-25 °C) for 10 min.
• Add 300 μl (2 amounts of sample volume) Binding Buffer and mix well by gently vortexing.

1b Preparation from nasal or throat swabs

• Transfer 150 μl PBS Buffer into a 1.5 ml microtube.
• Add 300 μl of Lysis Buffer.
• Cut off the cutton tip with the collected nasal or throat cells and place it in the micro tube.
• Close the tube and vortex for 15 sec.
• Incubate at room temperature (20-25 °C) for 10 min.
• Add 300 μl Binding Buffer and mix well by gently vortexing.

2 Column Loading

• Place a Spin Column into a provided 2 ml collection tube.
• Load lysate on the column and centrifuge at 13,000 g for 1 min.
• Note: The maximum volume of the column reservoirs 800 μl. For larger sample volumes discard the flow-through and load the column / spin again.
• Discard the flow-through in the collection tube and place the column back in the same tube.

3 Column Washing

• Add 500 μl Washing Buffer A to the Spin Column and centrifuge at 13,000 g for 1 min.
• Discard the flow-through in the collection tube and place the Spin Column back in the same tube.
• Add 500 μl Washing Buffer B to the Spin Column and centrifuge at 13,000 g for 1 min.
• Note: Before using Washing Buffer B for the first time add ethanol as indicated on the bottle.
• Discard the flow-through in the collection tube and place the Spin Column back in the same tube.
• Centrifuge at 13,000 g for 1 min.
• Note: It is important to dry the membrane since residual ethanol may interfere with downstream reactions.

4 Elution

• Place the Spin Column into a new 1.5 ml microtube (not provided).
• Add 30-60 μl Elution Buffer directly onto the membrane of the spin column.
• Note: Avoid touching membrane with the pipet tip.
• Incubate at room temperature for 1 min.
• Centrifuge at 13,000 g for 1 min.
• The eluted RNA and/or DNA is ready for down-stream processing. Use 2-5 μl as template for PCR or RT-PCR.

Product Citations:
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