Researcher: Can the amount of mRNA isolated using the mRNA Isolation Kits be increased by reusing the particles?

TechTALKer: The oligo (dT)₂₅-bound MPG^® Streptavidin magnetic solid support has a mRNA binding capacity of approximately 4 µg of mRNA per mg of particles. Optimal binding is achieved when isolating mRNA with 1 mg of MPG^® Streptavidin-oligo (dT)₂₅ from 0.05-0.1 g tissue, 10⁶-10⁷ cells or 150-300 µg total RNA. After eluting the mRNA, the particles can be washed and reused for up to two more rounds of isolation from the same tissue homogenate, cell lysate or total RNA prep, thus increasing the yield obtained per mg of particles. It is not recommended to regenerate the particles for use with different homogenates because you can never be sure that the mRNA from the previous isolation is completely removed. Although it is possible to strip the particles of residual mRNA, this may compromise the binding efficiency in subsequent purifications.

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