Cell and tissue lysis hub - Revision history

Jennifer Coalson: synonom addition, typo

2013-06-18T16:44:46Z

synonom addition, typo

←Older revision		Revision as of 16:44, 18 June 2013
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	=== Tissue lysis in liquid nitrogen ===		=== Tissue lysis in liquid nitrogen ===
		+	* also known as cryo-pulverization
	* best for tissue fragments (e.g. biopsy samples, tumor samples) or plant samples		* best for tissue fragments (e.g. biopsy samples, tumor samples) or plant samples
	* involves chilling the sample in liquid nitrogen and immediately afterwards subjecting to impact or grinding		* involves chilling the sample in liquid nitrogen and immediately afterwards subjecting to impact or grinding
-	* low cost, manual methods require a mortar and pestle or specialized stainless steel devices known as tissue ~~pulverizer~~	+	* low cost, manual methods require a mortar and pestle or specialized stainless steel devices known as tissue pulverizers
	* higher cost, mechanised methods involve a variety of specialized cryogenic cell disruption machines		* higher cost, mechanised methods involve a variety of specialized cryogenic cell disruption machines
	* plus: good protein extraction from small, tough samples		* plus: good protein extraction from small, tough samples

2013-06-18T16:42:13Z

new method, pros and cons

←Older revision		Revision as of 16:42, 18 June 2013
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	* 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		material: drills (polytron), pestle/tube (dounce) bead beaters
		+
		+	=== Tissue lysis in liquid nitrogen ===
		+	* best for tissue fragments (e.g. biopsy samples, tumor samples) or plant samples
		+	* involves chilling the sample in liquid nitrogen and immediately afterwards subjecting to impact or grinding
		+	* low cost, manual methods require a mortar and pestle or specialized stainless steel devices known as tissue pulverizer
		+	* higher cost, mechanised methods involve a variety of specialized cryogenic cell disruption machines
		+	* plus: good protein extraction from small, tough samples
		+	* negative: using some devices causes residual contamination between samples, not suitable for PCR amplification

	=== Freeze-thaw ===		=== Freeze-thaw ===

2008-12-19T10:45:41Z

Comparison of lysis methods

←Older revision		Revision as of 10:45, 19 December 2008
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	* 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 or bead beaters	+	material: drills (polytron), pestle/tube (dounce) bead beaters

	=== Freeze-thaw ===		=== Freeze-thaw ===

2008-09-24T15:43:36Z

material

←Older revision		Revision as of 15:43, 24 September 2008
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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 or 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 ==

2008-09-24T11:49:05Z

icon border removed

←Older revision		Revision as of 11:49, 24 September 2008
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-	[[Image:Hub icon.png\|right~~\|thumbnail\|'''protocol hub'''~~]]	+	[[Image:Hub icon.png\|right]]

	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.		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.

2008-09-24T11:47:52Z

lead

←Older revision		Revision as of 11:47, 24 September 2008
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	[[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. 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 ==

2008-06-27T13:12:25Z

Specific protocols

←Older revision		Revision as of 13:12, 27 June 2008
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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]]
Line 64:		Line 64:
	\| protein		\| protein
	\| detergent (SDS, NP40)		\| detergent (SDS, NP40)
-	\| ~~cell line~~	+	\| cells
	\|-		\|-
	\| [[Eccles:Protein Lysates from Tissue]]		\| [[Eccles:Protein Lysates from Tissue]]
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	\| RNA		\| RNA
	\| phenol		\| phenol
-	\| ~~cell line~~	+	\| cells
	\|-		\|-
	\| [[Sauer:RNA Purification from E. coli]]		\| [[Sauer:RNA Purification from E. coli]]

2008-06-26T11:56:18Z

Is this good enough?

←Older revision		Revision as of 11:56, 26 June 2008
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	== Specific protocols ==		== Specific protocols ==

-	{\| {{~~table~~}}	+	{\| {{sorttable}}
	! style="background:lightgrey"\|description/link		! style="background:lightgrey"\|description/link
	! style="background:lightgrey"\|target		! style="background:lightgrey"\|target

2008-06-26T11:10:03Z

Specific protocols

←Older revision		Revision as of 11:10, 26 June 2008
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	\| phenol		\| phenol
	\| cell line		\| cell line
		+	\|-
		+	\| [[Sauer:RNA Purification from E. coli]]
		+	\| RNA
		+	\| various options
		+	\| bacteria
	\|-		\|-
	\| add another method's name		\| add another method's name

2008-06-26T11:08:04Z