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Nucleic Acid Purification

GeneJET™ Viral DNA and RNA Purification Kit

Product information

Overview of purification procedure

Features

• Efficient purification of viral DNA and RNA from human and mammalian plasma, serum, blood and cell-free samples.
• Linear yields from a wide range of viral titers.
• Tested with a variety of DNA and RNA viruses (HIV, HBV, HCV, HPV, CMV, EBV).
• Downstream compatibility with (RT)-PCR, (RT)-qPCR and other nucleic acid based assays.

Applications

• Downstream use in qualitative and quantitative nucleic acid based assays, e.g. (RT)-PCR, (RT)-qPCR.

Description

The Thermo Scientific GeneJET Viral DNA and RNA Purification Kit is designed for rapid and efficient purification of high quality viral nucleic acids from various human and mammalian liquid samples such as plasma, serum, whole blood, saliva, nasal and buccal swabs, urine, urogenital swabs and milk. This kit is compatible with common sample preservatives such as EDTA, citrate and heparin.
The purified viral nucleic acids are free of proteins, nucleases, and other contaminants or inhibitors of downstream applications. Isolated DNA and RNA can be directly used in PCR, qPCR or other nucleic acid based assays.

Principle

The kit utilizes a silica-based membrane technology in the form of a convenient spin column. Viral nucleic acids from lysed samples bind to the column membrane while impurities are effectively removed during subsequent washing and centrifugation steps. Ready-to-use nucleic acids are eluted from the column.

Quality Control

Quality of the kit was tested by purification of RNA and DNA from 200 µl of spiked human plasma. Yield of nucleic acids was evaluated by RT-qPCR and qPCR, and compared with spiked copy number.

Components

• Column Preparation Liquid
• Lysis Solution
• Wash Buffer 1 (concentrated)
• Wash Buffer 2 (concentrated)
• Eluent
• Proteinase K
• Carrier RNA
• Spin Columns preassembled with Wash Tubes
• Wash Tubes (2 ml)

Storage

When the kit is delivered, remove the Carrier RNA from the package and store in the original aluminum bag at −20°C. Other components of the kit should be stored at room temperature (15-25°C).

Linear performance across a wide viral nucleic acid concentration range
Real-time PCR detection of Viral RNA (A) and DNA (B) purified with the GeneJET Viral DNA and RNA Purification Kit. Samples of various HBV and HCV titers were produced by diluting infected plasma of known viral titer with non-infected plasma.
A: Detection of RNA from various HCV sample titers.
B: Detection of DNA from various HBV sample titers.

High viral nucleic acid purification efficiency for various types of viruses
Different amounts of HBV, HCV, CMV and EBV infected human plasma or blood samples were purified using GeneJET Viral DNA and RNA Purification kit, and using kits from other suppliers. Isolated viral nucleic acids were quantified using a real-time PCR detection assay.

Reproducible results at low and high viral titers
Isolation of viral nucleic acids from multiple replicates of two plasma samples infected with high and low HCV (A) or HBV (B) titers using the GeneJET Viral DNA and RNA Purification Kit. Isolated HCV and HBV nucleic acids were quantified using a real-time PCR detection assay.
A: Detection of RNA from multiple replicates of HCV plasma samples with high and low viral titers.
B: Detection of DNA from multiple replicates of HBV plasma samples with high and low viral titers.

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Protocols & recommendations

RECOMMENDATIONS FOR USE