Restriction Digest: Partial - Revision history

Michael A. Speer: Partial Restriction Digest moved to Restriction Digest: Partial: To consolidate the digests on the menu

2009-06-18T12:18:33Z

Partial Restriction Digest moved to Restriction Digest: Partial: To consolidate the digests on the menu

←Older revision

Revision as of 12:18, 18 June 2009

Michael A. Speer at 12:17, 18 June 2009

2009-06-18T12:17:06Z

←Older revision	Revision as of 12:17, 18 June 2009
Line 1:	Line 1:
	+	==Abstract==
	+	This protocol can be used to generate genomic DNA fragments of different sizes using a common cutter. This is useful for many screening/probing processes.

	+	==Materials==
	+	* 10 microcentrifuge tubes
	+
	+	===Reagents===
	+	* Restriction Endonuclease
	+	* 10X Restriction Endonuclease buffer
	+	* BSA (10 mg/mL)
	+	* Genomic DNA (Concentration > 0.1 μg/μL)
	+
	+	==Procedure==
	+	# Label the microcentrifuge tubes 1 - 10
	+	# Mix together the following and invert to mix:
	+	## 180μL Genomic DNA
	+	## 20μL 10X Buffer
	+	## 2μL BSA
	+	# Dispense 40μL of this mix into tube 1 and 20μL into tubes 2 - 10 **[[Image:Critical step.png]]'''From this point keep everything on ice!!!'''
	+	# Add 1μL of restriction-endonuclease to tube 1 and invert to mix.
	+	# Transfer 20μL from tube 1 to tube 2 and invert to mix.
	+	# Repeat the last step transfering 20μL from the preceding tube to the succeeding tube mixing with every transfer.
	+	# Finally remove 20μL from tube 10
	+	# Incubate all 10 tubes for 1 hour at 37°C.
	+	# Finish the digest by submerging all 10 tubes in an 70°C water bath for 20 minutes.
	+	#Run the samples on a gel and choose the size which best fits your application.
	+
	+	==Acknowledgments==
	+	Acnkowledge any help you had in development, testing, writing this protocol.
	+
	+	==References==
	+	See [[OpenWetWare:Biblio]] for information on how to reference within a wiki.
	+
	+	==Specific Protocols==
	+	Add links to all the OWW protocols that have been used in making the consensus.
	+
	+	==Discussion==
	+	You can [[Talk:{{PAGENAME}}\|discuss this protocol]].
	+
	+
	+	[[Category:Protocol]]
	+	[[Category:In vitro]]
	+	[[Category:DNA]]

Michael A. Speer: Removing all content from page

2009-06-18T12:15:30Z

Removing all content from page

←Older revision		Revision as of 12:15, 18 June 2009
Line 1:		Line 1:
-	~~==Abstract==~~
-	~~This protocol can be used to generate genomic DNA fragments of different sizes using a common cutter. This is useful for many screening/probing processes.~~

-	~~==Materials==~~
-	* 10 microcentrifuge tubes
-
-	~~===Reagents===~~
-	* Restriction Endonuclease
-	* 10X Restriction Endonuclease buffer
-	* BSA (10 mg/mL)
-	* Genomic DNA (Concentration > 0.1 μg/μL)
-
-	~~==Procedure==~~
-	~~# Label the microcentrifuge tubes 1 - 10~~
-	~~# Mix together the following and invert to mix:~~
-	~~## 180μL Genomic DNA~~
-	~~## 20μL 10X Buffer~~
-	~~## 2μL BSA~~
-	~~# Dispense 40μL of this mix into tube 1 and 20μL into tubes 2 - 10 **[[Image:Critical step.png]]'''From this point keep everything on ice!!!'''~~
-	~~# Add 1μL of restriction-endonuclease to tube 1 and invert to mix.~~
-	~~# Transfer 20μL from tube 1 to tube 2 and invert to mix.~~
-	~~# Repeat the last step transfering 20μL from the preceding tube to the succeeding tube mixing with every transfer.~~
-	~~# Finally remove 20μL from tube 10~~
-	~~# Incubate all 10 tubes for 1 hour at 37°C.~~
-	~~# Finish the digest by submerging all 10 tubes in an 70°C water bath for 20 minutes.~~
-	~~#Run the samples on a gel and choose the size which best fits your application.~~
-
-	~~==Acknowledgments==~~
-	~~Acnkowledge any help you had in development, testing, writing this protocol.~~
-
-	~~==References==~~
-	~~See [[OpenWetWare:Biblio]] for information on how to reference within a wiki.~~
-
-	~~==Specific Protocols==~~
-	~~Add links to all the OWW protocols that have been used in making the consensus.~~
-
-	~~==Discussion==~~
-	~~You can [[Talk:{{PAGENAME}}\|discuss this protocol]].~~
-
-
-	~~[[Category:Protocol]]~~
-	~~[[Category:In vitro]]~~
-	~~[[Category:DNA]]~~

Michael A. Speer at 17:21, 11 June 2009

2009-06-11T17:21:02Z

←Older revision		Revision as of 17:21, 11 June 2009
Line 38:		Line 38:
	You can [[Talk:{{PAGENAME}}\|discuss this protocol]].		You can [[Talk:{{PAGENAME}}\|discuss this protocol]].

-	~~Tag this page with categories to allow easier indexing and searching. See [[Categories]] for information on existing categories.~~

	[[Category:Protocol]]		[[Category:Protocol]]
		+	[[Category:In vitro]]
		+	[[Category:DNA]]

Michael A. Speer: /* Procedure */

2009-06-11T17:19:53Z

Procedure

←Older revision		Revision as of 17:19, 11 June 2009
Line 17:		Line 17:
	## 20μL 10X Buffer		## 20μL 10X Buffer
	## 2μL BSA		## 2μL BSA
-	# Dispense 40μL of this mix into tube 1 and 20μL into tubes 2 - 10	+	# Dispense 40μL of this mix into tube 1 and 20μL into tubes 2 - 10 **[[Image:Critical step.png]]'''From this point keep everything on ice!!!'''
-	**[[Image:Critical step.png]]'''From this point keep everything on ice!!!'''	+
	# Add 1μL of restriction-endonuclease to tube 1 and invert to mix.		# Add 1μL of restriction-endonuclease to tube 1 and invert to mix.
	# Transfer 20μL from tube 1 to tube 2 and invert to mix.		# Transfer 20μL from tube 1 to tube 2 and invert to mix.
Line 26:		Line 25:
	# Finish the digest by submerging all 10 tubes in an 70°C water bath for 20 minutes.		# Finish the digest by submerging all 10 tubes in an 70°C water bath for 20 minutes.
	#Run the samples on a gel and choose the size which best fits your application.		#Run the samples on a gel and choose the size which best fits your application.
-

	==Acknowledgments==		==Acknowledgments==

Michael A. Speer: New page: ==Abstract== This protocol can be used to generate genomic DNA fragments of different sizes using a common cutter. This is useful for many screening/probing processes. ==Materials== * ...

2009-06-11T17:19:26Z

New page: ==Abstract== This protocol can be used to generate genomic DNA fragments of different sizes using a common cutter. This is useful for many screening/probing processes. ==Materials== * ...

New page

==Abstract==
This protocol can be used to generate genomic DNA fragments of different sizes using a common cutter. This is useful for many screening/probing processes.

==Materials==
* 10 microcentrifuge tubes

===Reagents===
* Restriction Endonuclease
* 10X Restriction Endonuclease buffer
* BSA (10 mg/mL)
* Genomic DNA (Concentration > 0.1 μg/μL)

==Procedure==
# Label the microcentrifuge tubes 1 - 10
# Mix together the following and invert to mix:
## 180μL Genomic DNA
## 20μL 10X Buffer
## 2μL BSA
# Dispense 40μL of this mix into tube 1 and 20μL into tubes 2 - 10
**[[Image:Critical step.png]]'''From this point keep everything on ice!!!'''
# Add 1μL of restriction-endonuclease to tube 1 and invert to mix.
# Transfer 20μL from tube 1 to tube 2 and invert to mix.
# Repeat the last step transfering 20μL from the preceding tube to the succeeding tube mixing with every transfer.
# Finally remove 20μL from tube 10
# Incubate all 10 tubes for 1 hour at 37°C.
# Finish the digest by submerging all 10 tubes in an 70°C water bath for 20 minutes.
#Run the samples on a gel and choose the size which best fits your application.

==Acknowledgments==
Acnkowledge any help you had in development, testing, writing this protocol.

==References==
See [[OpenWetWare:Biblio]] for information on how to reference within a wiki.

==Specific Protocols==
Add links to all the OWW protocols that have been used in making the consensus.

==Discussion==
You can [[Talk:{{PAGENAME}}|discuss this protocol]].

Tag this page with categories to allow easier indexing and searching. See [[Categories]] for information on existing categories.

[[Category:Protocol]]