Cell and tissue lysis hub

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This page should point you to the many different general and lab-specific protocols describing tissue and cell lysis and serve as forum for comparison. The ups and downs of various methods from sonication, homogenization, freeze-thaw cycles, and detergent-based lysis are examined below.

Comparison of lysis methods

Sonication

  • most efficient method of cell fractionation
  • problem: heat build up which can denature proteins (proportionate to length of sonication)
  • precaution: do on ice and sonicate intermittantly

material: ultrasonication bath or rods

Homogenisation

  • best for animal tissue; less suitable for cells
  • precaution: do on ice to reduce heat build-up and denaturation

material: drills (polytron), pestle/tube (dounce) bead beaters

Tissue lysis in liquid nitrogen

  • also known as cryo-pulverization
  • best for tissue fragments (e.g. biopsy samples, tumor samples) or plant samples
  • involves chilling the sample in liquid nitrogen and immediately afterwards subjecting to impact or grinding
  • low cost, manual methods require a mortar and pestle or specialized stainless steel devices known as tissue pulverizers
  • higher cost, mechanised methods involve a variety of specialized cryogenic cell disruption machines
  • plus: good protein extraction from small, tough samples
  • negative: using some devices causes residual contamination between samples, not suitable for PCR amplification

Freeze-thaw

  • least effective method
  • plus: does not denature proteins as much as other methods

material: pestle and mortar, liquid nitrogen

Detergents

  • chemical method of lysis
  • problems: detergent may inhibit subsequent reactions
  • problems: detergent may disrupts protein interactions

no additional material required, just the chemicals and typical tubes

Specific protocols

description/link target lysis method type of material
Silver: Lysate for Western protein detergent (Triton) cells
Sauer:Lysing E. coli with Lysozymes lysozyme bacteria
Blackburn:Yeast Colony PCR DNA NaOH yeast
Jacobs:Protocol Total Protein Isolation Using RIPA Lysis Buffer protein detergent (RIPA buffer) cells
Streptomyces:Protocols/Mini-Maxi Prep DNA NaOH bacteria
Western Blot/Tissue Preparation protein detergent & physical disruption tissue
Eccles:Protein Lysates from Cells in Culture protein detergent (SDS, NP40) cells
Eccles:Protein Lysates from Tissue protein detergent, physical, freezing tissue
RNA extraction using trizol/tri RNA phenol cells
Sauer:RNA Purification from E. coli RNA various options bacteria
add another method's name target lysis method type of starting material

Related hub pages

Related discussions on BioForum