Isolation and Visualization of Rotavirus Genome via Gel Electrophoresis

Take a debris-free cell lysate containing rotavirus.
Add a denaturing salt solution and vortex. The salt denatures host proteins, inactivates RNases, and destabilizes the viral capsid.
Add chloroform and vortex to rupture the viral capsid, releasing viral RNA segments.
Centrifuge the suspension. The RNA collects in the aqueous phase, while denatured capsid proteins accumulate at the interface.
Transfer the aqueous phase and add isopropyl alcohol, which lowers nucleic acid solubility and precipitates RNA.
Centrifuge to separate RNA. Discard the supernatant, and add ethanol.
Centrifuge to pellet RNA and discard the supernatant.
Air-dry to remove traces of ethanol, then dissolve RNA in nuclease-free water to preserve RNA integrity.
Mix RNA with loading buffer and load it onto a polyacrylamide gel.
Run electrophoresis for size-based separation of viral RNA segments.
Stain the RNA with an RNA-intercalating dye.
Under ultraviolet light, eleven distinct bands confirm recovery of the rotavirus genome.
Add 600 microliters of clarified infected cell lysates and 400 microliters of guanidinium thiocyanate into 1.5 milliliter microcentrifuge tubes. Vortex them for 30 seconds and incubate them at room tempe