Research use only. Not for use in diagnostic procedures. This protocol is relevant for catalog number RNB100.
The GenElute™ Total RNA Purification Kit provides a rapid method for the isolation and purification of total RNA from viruses such as the 2019 Novel Coronavirus (SARS-CoV-2), which causes COVID-19 disease. Purification is based on spin column chromatography using a proprietary resin as the separation matrix. The RNA is preferentially purified from other cellular components, such as proteins, without the use of phenol or chloroform. The procedure involves:
The protocol is designed to yield purified RNA of the highest integrity that can be used in various downstream applications including PCR, Northern blotting, and expression array assays.
GenElute™ Viral RNA Purification Protocol:
Note: Ensure that all solutions are at room temperature prior to use. The maximum recommended number of cells is 3 x 106. Cell pellets can be stored at –70 °C for later use or used directly in the procedure. Determine the number of cells present before freezing. Frozen pellets should be stored for no longer than 2 weeks to ensure that the integrity of the RNA is not compromised. Frozen cell pellets should not be thawed prior to beginning the protocol. Add the Buffer RL directly to the frozen cell pellet.
1 For starting amounts greater than 106 cells, it is recommended that the lysate is passed through a 25 gauge needle attached to a syringe 5-10 times at this point, in order to shear the genomic DNA prior to loading onto the column.
2 For starting amounts greater than 106 cells, it is recommended that the lysate is passed through a 25 gauge needle attached to a syringe 5-10 times at this point, in order to shear the genomic DNA prior to loading onto the column.
Tissue lysis procedure:
Notes: This procedure is for the isolation of RNA from whole blood. It is recommended that no more than 100 µL of blood be used in order to prevent clogging of the column. We recommend the use of this kit to isolate RNA from non-coagulating fresh blood using EDTA as the anti-coagulant. Working quickly is important for this procedure.
Whole blood lysis procedure:
Plasma and serum lysis procedure:
Notes: Body fluid of all human and animal subjects is considered potentially infectious. It is important to work quickly during this procedure.
Nasal or throat swab lysis procedure:
Free viral particle lysis procedure:
Binding RNA to Column
The GenElute™ Total RNA Purification Kit isolates total RNA with minimal amounts of genomic DNA contamination. As an optional step, the On-Column DNA Removal Protocol can be performed at this point for maximum removal of residual DNA that may affect sensitive downstream applications. Please refer to separate instructions provided in the user guide to perform the On-Column DNA Removal using Catalog Number DNASE10-1SET (not included in kit).
The purified RNA sample may be stored at –20 °C for a few days. It is recommended that samples be placed at –70 °C for long term storage.