Cell and tissue lysis hub: Difference between revisions
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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. | 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. | ||
== Comparison of lysis methods == | == Comparison of lysis methods == | ||
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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 | ||
== Specific protocols == | |||
* [[Silver: Lysate for Western]] - Silver Lab: cell lysis for [[Western]] (method: Triton) | |||
* [[Sauer:Lysing E. coli with Lysozymes]] - Sauer Lab: bacteria lysis (method: lysozyme) | |||
* [[Blackburn:Yeast Colony PCR]] - Blackburn Lab: yeast lysis for [[colony PCR]] (method NaOH) | |||
* [[Jacobs:Protocol Total Protein Isolation Using RIPA Lysis Buffer]] - Jacobs Lab: cell line lysis for protein (method: RIPA buffer) | |||
* [[Streptomyces:Protocols/Mini-Maxi Prep]] - Streptomyces community, East Anglia: bacteria lysis for DNA (method: NaOH) | |||
Revision as of 03:17, 26 June 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.
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
Homogenisation
- best for animal tissue; less suitable for cells
- precaution: do on ice to reduce heat build-up and denaturation
Freeze-thaw
- least effective method
- plus: does not denature proteins as much as other methods
Detergents
- chemical method of lysis
- problems: detergent may inhibit subsequent reactions
- problems: detergent may disrupts protein interactions
Specific protocols
- Silver: Lysate for Western - Silver Lab: cell lysis for Western (method: Triton)
- Sauer:Lysing E. coli with Lysozymes - Sauer Lab: bacteria lysis (method: lysozyme)
- Blackburn:Yeast Colony PCR - Blackburn Lab: yeast lysis for colony PCR (method NaOH)
- Jacobs:Protocol Total Protein Isolation Using RIPA Lysis Buffer - Jacobs Lab: cell line lysis for protein (method: RIPA buffer)
- Streptomyces:Protocols/Mini-Maxi Prep - Streptomyces community, East Anglia: bacteria lysis for DNA (method: NaOH)
