RNEASY KITS User Manual

Loading ...

RNeasy Mini Handbook 10/2019

Table 3. Disruption and homogenization methods

* QIAshredder homogenizers are supplied in the RNeasy Plant Mini Kit and can be purchased separately for

use with the RNeasy Mini Kit or RNeasy Protect Mini Kit. See page 91 for ordering information.

Disruption and homogenization using the TissueLyser system

In bead-milling, cells and tissues can be disrupted by rapid agitation in the presence of beads

and lysis buffer. Disruption and simultaneous homogenization occur by the shearing and crushing

action of the beads as they collide with the cells. Disruption efficiency is influenced by:

 Size and composition of beads

 Ratio of buffer to beads

 Amount of starting material

 Speed and configuration of the TissueLyser

 Disintegration time

Sample Disruption method Homogenization method Comments

Animal cells Addition of lysis buffer Rotor–stator homogenizer or

QIAshredder homogenizer*

or syringe and needle

If processing ≤1 x 10

cells,

lysate can be homogenized by

vortexing

Animal tissues TissueLyser TissueLyser

The TissueLyser gives results

comparable to using a rotor–

stator homogenizer

Rotor–stator

homogenizer

Rotor–stator homogenizer Simultaneously disrupts and

homogenizes

Mortar and pestle

QIAshredder homogenizer*

or syringe and needle

Rotor–stator homogenizer usually

gives higher yields than mortar

and pestle

Yeast

Enzymatic digestion of

cell wall followed by

lysis of spheroplasts

Vortexing

TissueLyser with glass

beads

TissueLyser with glass beads

TissueLyser simultaneously disrupts

and homogenizes; cannot be

replaced by vortexing

Plants and

filamentous fungi

Mortar and pestle QIAshredder homogenizer*

Mortar and pestle cannot be

replaced by rotor–stator

homogenizer

Loading ...

User Manual - Page 23