Cell and tissue lysis hub

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(comparison of lysis method section)
(Comparison of lysis methods)
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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.
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* [[Silver: Lysate for Western]] - Silver Lab: cell lysis for [[Western]] (method: Triton)
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* [[Sauer:Lysing E. coli with Lysozymes]] - Sauer Lab: bacteria lysis (method: lysozyme)
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* [[Blackburn:Yeast Colony PCR]] - Blackburn Lab: yeast lysis for [[colony PCR]] (method NaOH)
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* [[Jacobs:Protocol Total Protein Isolation Using RIPA Lysis Buffer]] - Jacobs Lab: cell line lysis for protein (method: RIPA buffer)
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* [[Streptomyces:Protocols/Mini-Maxi Prep]] - Streptomyces community, East Anglia: bacteria lysis for DNA (method: NaOH)
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== 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
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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
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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
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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
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no additional material required, just the chemicals and typical tubes
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== Specific protocols ==
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{| {{sorttable}}
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! style="background:lightgrey"|description/link
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! style="background:lightgrey"|target
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! style="background:lightgrey"|lysis method
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! style="background:lightgrey"|type of material
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|-
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| [[Silver: Lysate for Western]]
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| protein
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| detergent (Triton)
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| cells
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|-
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| [[Sauer:Lysing E. coli with Lysozymes]]
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|
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| lysozyme
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| bacteria
 +
|-
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| [[Blackburn:Yeast Colony PCR]]
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| DNA
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| NaOH
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| yeast
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|-
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| [[Jacobs:Protocol Total Protein Isolation Using RIPA Lysis Buffer]]
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| protein
 +
| detergent (RIPA buffer)
 +
| cells
 +
|-
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| [[Streptomyces:Protocols/Mini-Maxi Prep]]
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| DNA
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| NaOH
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| bacteria
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|-
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| [[Western Blot/Tissue Preparation]]
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| protein
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| detergent & physical disruption
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| tissue
 +
|-
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| [[Eccles:Protein Lysates from Cells in Culture]]
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| protein
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| detergent (SDS, NP40)
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| cells
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|-
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| [[Eccles:Protein Lysates from Tissue]]
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| protein
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| detergent, physical, freezing
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| tissue
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|-
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| [[RNA extraction using trizol/tri]]
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| RNA
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| phenol
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| cells
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|-
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| [[Sauer:RNA Purification from E. coli]]
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| RNA
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| various options
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| bacteria
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|-
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| add another method's name
 +
| target
 +
| lysis method
 +
| type of starting material
 +
|}
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== Related hub pages ==
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* [[RNA extraction]]
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== Related discussions on BioForum ==
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* [http://www.protocol-online.org/forums/index.php?showtopic=23747 cell lysis methods comparison]
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* [http://www.protocol-online.org/forums/index.php?showtopic=1943&hl=cell+lysis what solution to use for cell lysis]
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[[Category:Protocol]][[Category:DNA]][[Category:RNA]][[Category:protein]]
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[[Category:Material]][[Category:Chemical]]
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[[Category:Escherichia coli]][[Category:Mouse]][[Category:Streptomyces]][[Category:Yeast]]

Revision as of 05:45, 19 December 2008

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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