Cell and tissue lysis hub: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
(several protocols into table) |
||
| Line 1: | Line 1: | ||
[[Image:Hub icon.png|right|thumbnail|protocol hub]] | [[Image:Hub icon.png|right|thumbnail|'''protocol hub''']] | ||
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. | ||
| Line 61: | Line 61: | ||
| tissue | | tissue | ||
|- | |- | ||
| | | [[Eccles:Protein Lysates from Cells in Culture]] | ||
| | | protein | ||
| | | detergent (SDS, NP40) | ||
| | | cell line | ||
|- | |||
| [[Eccles:Protein Lysates from Tissue]] | |||
| protein | |||
| detergent, physical, freezing | |||
| tissue | |||
|- | |- | ||
| | | [[RNA extraction using trizol/tri]] | ||
| | | RNA | ||
| | | phenol | ||
| | | cell line | ||
|- | |- | ||
| add another method's name | | add another method's name | ||
| Line 76: | Line 81: | ||
| type of starting material | | type of starting material | ||
|} | |} | ||
== Related hub pages == | == Related hub pages == | ||
Revision as of 04:07, 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 |
| Eccles:Protein Lysates from Cells in Culture | protein | detergent (SDS, NP40) | cell line |
| Eccles:Protein Lysates from Tissue | protein | detergent, physical, freezing | tissue |
| RNA extraction using trizol/tri | RNA | phenol | cell line |
| add another method's name | target | lysis method | type of starting material |
