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RNeasy Mini Handbook 10/2019
25
Disruption using a mortar and pestle
For disruption using a mortar and pestle, freeze the animal or plant tissue immediately in liquid
nitrogen and grind to a fine powder under liquid nitrogen. Transfer the suspension (tissue
powder and liquid nitrogen) into a liquid-nitrogen–cooled, appropriately sized tube and allow
the liquid nitrogen to evaporate without allowing the sample to thaw. Add lysis buffer and
continue as quickly as possible with the homogenization according to one of the 2 methods
below.
Note: Grinding the sample using a mortar and pestle will disrupt the sample, but will not
homogenize it. Homogenization must be performed afterwards.
Homogenization using QIAshredder homogenizers
Using QIAshredder homogenizers is a fast and efficient way to homogenize cell and tissue
lysates without cross-contamination of samples. Up to 700 µl of lysate is loaded onto a
QIAshredder spin column placed in a 2 ml collection tube, and spun for 2 min at maximum
speed in a microcentrifuge. The lysate is homogenized as it passes through the spin column.
QIAshredder spin columns are supplied in the RNeasy Plant Mini Kit and can be purchased
separately for use with the RNeasy Mini Kit and RNeasy Protect Mini Kit. See page 91 for
ordering information.
Homogenization using a syringe and needle
Cell and tissue lysates can be homogenized using a syringe and needle. Lysate is passed
through a 20-gauge (0.9 mm) needle attached to a sterile plastic syringe at least 5–10 times
or until a homogeneous lysate is achieved. Increasing the volume of lysis buffer may be
required to facilitate handling and minimize loss.
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