Cell and tissue lysis hub

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[[Image:Hub icon.png|right]]
[[Image:Hub icon.png|right]]
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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.
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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 ==
== Comparison of lysis methods ==
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* problem: heat build up which can denature proteins (proportionate to length of sonication)
* problem: heat build up which can denature proteins (proportionate to length of sonication)
* precaution: do on ice and sonicate intermittantly
* precaution: do on ice and sonicate intermittantly
 +
material: ultrasonication bath or rods
=== Homogenisation ===  
=== Homogenisation ===  
* best for animal tissue; less suitable for cells
* best for animal tissue; less suitable for cells
* precaution: do on ice to reduce heat build-up and denaturation
* precaution: do on ice to reduce heat build-up and denaturation
 +
material: drills (polytron), pestle/tube (dounce) bead beaters
=== Freeze-thaw ===
=== Freeze-thaw ===
* least effective method
* least effective method
* plus: does not denature proteins as much as other methods
* plus: does not denature proteins as much as other methods
 +
material: pestle and mortar, liquid nitrogen
=== Detergents ===
=== Detergents ===
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* problems: detergent may inhibit subsequent reactions
* problems: detergent may inhibit subsequent reactions
* problems: detergent may disrupts protein interactions
* problems: detergent may disrupts protein interactions
 +
no additional material required, just the chemicals and typical tubes
== Specific protocols ==
== Specific protocols ==
-
{| {{table}}
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{| {{sorttable}}
! style="background:lightgrey"|description/link
! style="background:lightgrey"|description/link
! style="background:lightgrey"|target
! style="background:lightgrey"|target
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| protein
| protein
| detergent (Triton)
| detergent (Triton)
-
| eukaryotic cells
+
| cells
|-
|-
| [[Sauer:Lysing E. coli with Lysozymes]]
| [[Sauer:Lysing E. coli with Lysozymes]]
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| protein
| protein
| detergent (RIPA buffer)
| detergent (RIPA buffer)
-
| cell line
+
| cells
|-
|-
| [[Streptomyces:Protocols/Mini-Maxi Prep]]
| [[Streptomyces:Protocols/Mini-Maxi Prep]]
-
| DAN
+
| DNA
| NaOH
| NaOH
| bacteria
| bacteria
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| detergent & physical disruption
| detergent & physical disruption
| tissue
| 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]]
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| RNA
 +
| phenol
 +
| cells
 +
|-
 +
| [[Sauer:RNA Purification from E. coli]]
 +
| RNA
 +
| various options
 +
| bacteria
|-
|-
| add another method's name
| add another method's name
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| type of starting material
| type of starting material
|}
|}
 +
 +
== Related hub pages ==
 +
* [[RNA extraction]]
== Related discussions on BioForum ==
== Related discussions on BioForum ==

Current revision

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.

Contents

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

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

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