Griffitts:Restriction digest - Revision history

Matthew B Biggs: /* Enzymes */

2012-03-20T18:38:32Z

Enzymes

2012-03-20T18:38:08Z

Enzymes

2009-04-13T16:49:11Z

Double Digests: added footnote

←Older revision		Revision as of 16:49, 13 April 2009
Line 295:		Line 295:
	\|-		\|-
	\| '''''Bam'' HI'''		\| '''''Bam'' HI'''
-	\| seq<sup>1</sup>	+	\| seq<sup><font color=red>1</font></sup>
	\| -		\| -
	\| -		\| -
Line 517:		Line 517:
	\| '''''Xba'' I'''		\| '''''Xba'' I'''
	\| 2		\| 2
-	\| seq	+	\| seq<sup><font color=red>2</font></sup>
	\| 2		\| 2
	\| 2		\| 2
Line 550:		Line 550:
	\|-		\|-
	\|}		\|}
-	<sup>1</sup> Sequential digest recommended.<br>	+	<sup><font color=red>1</font></sup>Sequential digest recommended.<br>
		+	<sup><font color=red>2</font></sup>We actually prefer to do the double-digest and use Buffer 2 for the ''Bam''HI–''Xba''I double-digest.<br>
	Note that a double-digest of ''Bam'' HI and ''Xba'' I is not recommended. However, by following [[Griffitts:Restriction digest#30 μL reaction\|the procedure above]] you won't have any problems.		Note that a double-digest of ''Bam'' HI and ''Xba'' I is not recommended. However, by following [[Griffitts:Restriction digest#30 μL reaction\|the procedure above]] you won't have any problems.
	<br><br>		<br><br>
-	''Adapted from New England BioLabs 2005-06 Catalog and Technical Reference.''	+	Adapted from ''New England BioLabs 2005-06 Catalog and Technical Reference.''

2009-04-13T16:44:46Z

Recipes

←Older revision		Revision as of 16:44, 13 April 2009
Line 15:		Line 15:
	* 5 μL DNA		* 5 μL DNA
	* 1.5 μL appropriate 10X buffer		* 1.5 μL appropriate 10X buffer
-	* 1.5 μL 10X BSA (if needed--see below)	+	* 1.5 μL 10X BSA (if needed—see below)
	* Restriction enzyme(s) (see specific enzymes below for exact amounts)		* Restriction enzyme(s) (see specific enzymes below for exact amounts)
	** Don't immerse the tip, draw from the surface to avoid excess enzyme		** Don't immerse the tip, draw from the surface to avoid excess enzyme
Line 25:		Line 25:
	* 12 μL DNA		* 12 μL DNA
	* 3.0 μL appropriate 10X buffer		* 3.0 μL appropriate 10X buffer
-	* 3.0 μL 10X BSA (if needed--see below)	+	* 3.0 μL 10X BSA (if needed—see below)
	* Restriction enzyme(s) (see specific enzymes below for exact amounts)		* Restriction enzyme(s) (see specific enzymes below for exact amounts)
	** Don't immerse the tip, draw from the surface to avoid excess enzyme		** Don't immerse the tip, draw from the surface to avoid excess enzyme

2009-04-13T16:44:25Z

40 μL reaction

←Older revision		Revision as of 16:44, 13 April 2009
Line 34:		Line 34:
	===40 μL reaction===		===40 μL reaction===
	* 20 μL DNA		* 20 μL DNA
-	* 4.3 μL appropriate 10X buffer	+	* 4.0 μL appropriate 10X buffer
-	* 4.0 μL 10X BSA (if needed--see below)	+	* 4.0 μL 10X BSA (if needed—see below)
	* Restriction enzyme(s) (see specific enzymes below for exact amounts)		* Restriction enzyme(s) (see specific enzymes below for exact amounts)
	** Don't immerse the tip, draw from the surface to avoid excess enzyme		** Don't immerse the tip, draw from the surface to avoid excess enzyme
	* Add ddH<sub>2</sub>0 to bring the final volume to 40 μL		* Add ddH<sub>2</sub>0 to bring the final volume to 40 μL
-	Note: Use this recipe when your DNA concentrations are low	+	Note: Use this recipe when your DNA concentrations are low<br>
		+	Note: For a ''Bam''HI–''Xba''I double-digest, use 4.3 μL of 10X buffer

	==Procedure==		==Procedure==

2009-04-13T16:43:54Z

30 μL reaction

←Older revision		Revision as of 16:43, 13 April 2009
Line 29:		Line 29:
	** Don't immerse the tip, draw from the surface to avoid excess enzyme		** Don't immerse the tip, draw from the surface to avoid excess enzyme
	* Add ddH<sub>2</sub>0 to bring the final volume to 30 μL		* Add ddH<sub>2</sub>0 to bring the final volume to 30 μL
-	Note: Use this recipe when your DNA concentrations are sufficient	+	Note: Use this recipe when your DNA concentrations are sufficient<br>
	Note: For a ''Bam''HI–''Xba''I double-digest, use 3.3 μL of 10X buffer		Note: For a ''Bam''HI–''Xba''I double-digest, use 3.3 μL of 10X buffer

2009-04-13T16:43:22Z

30 μL reaction

←Older revision		Revision as of 16:43, 13 April 2009
Line 24:		Line 24:
	===30 μL reaction===		===30 μL reaction===
	* 12 μL DNA		* 12 μL DNA
-	* 3.3 μL appropriate 10X buffer	+	* 3.0 μL appropriate 10X buffer
	* 3.0 μL 10X BSA (if needed--see below)		* 3.0 μL 10X BSA (if needed--see below)
	* Restriction enzyme(s) (see specific enzymes below for exact amounts)		* Restriction enzyme(s) (see specific enzymes below for exact amounts)
Line 30:		Line 30:
	* Add ddH<sub>2</sub>0 to bring the final volume to 30 μL		* Add ddH<sub>2</sub>0 to bring the final volume to 30 μL
	Note: Use this recipe when your DNA concentrations are sufficient		Note: Use this recipe when your DNA concentrations are sufficient
		+	Note: For a ''Bam''HI–''Xba''I double-digest, use 3.3 μL of 10X buffer

	===40 μL reaction===		===40 μL reaction===

2009-04-13T16:42:20Z

15 μL reaction

←Older revision		Revision as of 16:42, 13 April 2009
Line 14:		Line 14:
	===15 μL reaction===		===15 μL reaction===
	* 5 μL DNA		* 5 μL DNA
-	* 1.8 μL appropriate 10X buffer	+	* 1.5 μL appropriate 10X buffer
	* 1.5 μL 10X BSA (if needed--see below)		* 1.5 μL 10X BSA (if needed--see below)
	* Restriction enzyme(s) (see specific enzymes below for exact amounts)		* Restriction enzyme(s) (see specific enzymes below for exact amounts)

2008-04-11T17:08:22Z

←Older revision		Revision as of 17:08, 11 April 2008
Line 1:		Line 1:
-	{{Griffitts}}	+	<div class="noprint">
		+	{{Template:Griffitts}}
		+	</div>
	<font size="5">Restriction Digest</font>		<font size="5">Restriction Digest</font>
	==Materials==		==Materials==

2008-01-17T16:48:40Z

Double Digests

←Older revision		Revision as of 16:48, 17 January 2008
Line 547:		Line 547:
	\|}		\|}
	<sup>1</sup> Sequential digest recommended.<br>		<sup>1</sup> Sequential digest recommended.<br>
-	Note that a double-digest of ''Bam'' HI and ''Xba'' I is not recommended. By following [[Griffitts:Restriction digest#30 μL reaction\|the procedure above]] you won't have any problems.	+	Note that a double-digest of ''Bam'' HI and ''Xba'' I is not recommended. However, by following [[Griffitts:Restriction digest#30 μL reaction\|the procedure above]] you won't have any problems.
	<br><br>		<br><br>
	''Adapted from New England BioLabs 2005-06 Catalog and Technical Reference.''		''Adapted from New England BioLabs 2005-06 Catalog and Technical Reference.''