Description
The EZBioscience® EZ-press Viral RNA Purification Kit provides a simple, rapid and efficient method to simultaneously purify viral RNA from fresh or frozen cell-free biological fluids (plasma, serum, cerebrospinal fluid, etc.) and cell culture supernatants. The purified Viral RNA is suitable for use in a variety of downstream applications, including RT-PCR, qRT-PCR, and qPCR, and can be used for viral load monitoring, viral detection and genotyping. Cell-free samples are first lysed and homogenized in a strong denaturant and phenol containing buffer, which immediately inactivates RNase to ensure isolation of intact RNA. After homogenization, Buffer A is mixed with the lysate and incubated. And then the mixture is centrifuged for phase separation; the upper-layer supernatant is collected and mixed with Supplemental Reagent and ethanol. The liquid is then passed through the RNA binding Spin Column containing a silica-based membrane to which the RNA binds. Impurities are effectively removed by subsequent washing. The purified total RNA is then eluted with Elution Buffer.
Components
Components | EZB-VRN1 (100 Preps) |
Lysis Buffer | 55 ml |
Buffer A | 22 ml |
Supplemental Reagent | 1100 μl |
Wash Buffer 1* | 13 ml |
Wash Buffer 2* | 13 ml |
Elution Buffer | 10 ml |
Spin Columns for RNA (with Collection Tubes) | 100 Preps |
*: Before using for the first time, add 52 ml of 100% ethanol to the Wash Buffer 1, 52 ml of 100% ethanol to the Wash Buffer 2.
Storage
Store the Lysis Buffer and Buffer A at 2 ~ 8°C, protect from light. Store the Supplemental Reagent at -20°C. Store other components at room temperature (When using these buffers, be careful to avoid of contamination). Divide the Elution Buffer into small aliquots upon reception is suggested.
Precautions
1. Handle all viruses in compliance with established institutional guidelines. Since safety requirements for use and handling of viruses may vary at individual institutions, we recommend consulting the health and safety guidelines and/or officers at your institution.
2. Be sure to take the appropriate precautions (wear a laboratory coat, disposable gloves, and eye protection) when handling viral samples.
Experimental Procedure at a Glance
Features
1. Easy-to-follow protocol for lysis and homogenization.
2. The kit contains Buffer A, which can replace chloroform.
3. Isolation from fresh or frozen cell-free biological fluids (plasma, serum, cerebrospinal fluid, etc.) and cell culture supernatants.
4. High-purity RNA suitable for all downstream applications: RT-PCR, RT-qPCR, RNA Sequencing, and so on.
Representative Experimental Results
Figure1. RNA isolated from Lentivirus RNA sample using EZBioscience® EZ-press Viral RNA Purification Kit. The qPCR detection result shows that this kit can be well used for the purification of viral RNA.
Figure 2. The N gene of SARS-CoV-2 Virus expression level in equivalently diluted RNA purified from SARS-CoV-2 Pseudovirus were detected. The amplification plots above indicate that, the target gene could be amplified efficiently and specifically. And the linear relationship within the detection range is very good.
For detailed information, please check the product manual.
