Cell and tissue lysis hub: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
(→Specific protocols: protocol added) |
|||
Line 52: | Line 52: | ||
|- | |- | ||
| [[Streptomyces:Protocols/Mini-Maxi Prep]] | | [[Streptomyces:Protocols/Mini-Maxi Prep]] | ||
| | | DNA | ||
| NaOH | | NaOH | ||
| bacteria | | bacteria | ||
Line 65: | Line 65: | ||
| lysis method | | lysis method | ||
| type of starting material | | type of starting material | ||
|} | |- | ||
| add another method's name | |||
| target | |||
| lysis method | |||
| type of starting material|} | |||
|- | |||
| add another method's name | |||
| target | |||
| lysis method | |||
| type of starting material | |||
== Related discussions on BioForum == | == Related discussions on BioForum == |
Revision as of 03:57, 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
description/link | target | lysis method | type of material |
---|---|---|---|
Silver: Lysate for Western | protein | detergent (Triton) | eukaryotic 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) | cell line |
Streptomyces:Protocols/Mini-Maxi Prep | DNA | NaOH | bacteria |
Western Blot/Tissue Preparation | protein | detergent & physical disruption | tissue |
add another method's name | target | lysis method | type of starting material |
add another method's name | target | lysis method | } |
add another method's name | target | lysis method | type of starting material
Related discussions on BioForum |