Richard Lab:Agarose Gel Electrophoresis - Revision history

Michael A. Speer: /* Notes */

2009-06-23T18:50:52Z

Notes

←Older revision		Revision as of 18:50, 23 June 2009
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	[[Category:Protocol]] [[Category:In vitro]] [[Category:DNA]]		[[Category:Protocol]] [[Category:In vitro]] [[Category:DNA]]

-	Back to [[Richard_Lab \| ~~Richard Lab~~]]	+	Back to [[Richard_Lab:protocols \| Protocols]]

2009-06-23T18:50:20Z

Notes

←Older revision		Revision as of 18:50, 23 June 2009
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	** Correspondingly, the 10X TBE is officially 5X.		** Correspondingly, the 10X TBE is officially 5X.
	*Using TBE allows us to have such a high (150V) voltage. If you use TAE you need to use a significantly lower voltage and your run time will be longer. Despite that, TAE is advantagous in some cases, but [[User:Michael_A. Speer \| Mike]] feels that TBE is better suited for his applications. This is discussed in detail in the [[agarose gel electrophoresis \| consensus protocol]].		*Using TBE allows us to have such a high (150V) voltage. If you use TAE you need to use a significantly lower voltage and your run time will be longer. Despite that, TAE is advantagous in some cases, but [[User:Michael_A. Speer \| Mike]] feels that TBE is better suited for his applications. This is discussed in detail in the [[agarose gel electrophoresis \| consensus protocol]].
-	* Use the following table to determine the amount of agarose you want to use.	+	* Use the following table to determine the amount of agarose you want to use.<br><br>
	<center>		<center>
	{\| border="1" cellpadding="5" cellspacing="0" align="center"		{\| border="1" cellpadding="5" cellspacing="0" align="center"

2009-06-23T18:49:15Z

←Older revision

Revision as of 18:49, 23 June 2009

2009-06-23T17:58:02Z

Procedure

←Older revision		Revision as of 17:58, 23 June 2009
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	## 10ul DNA ladder		## 10ul DNA ladder
	## 1ul [[SYBR_Green_II\|SYBR green]] (100X)		## 1ul [[SYBR_Green_II\|SYBR green]] (100X)
-	## 1ul Bromophenol Blue	+	## 1ul [[Agarose_gel_loading_dye\|Bromophenol Blue]]
	# Prepare the DNA Samples by combining the following:		# Prepare the DNA Samples by combining the following:
	## 40ul DNA preparation		## 40ul DNA preparation

2009-06-23T17:54:52Z

Notes

←Older revision		Revision as of 17:54, 23 June 2009
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	===Notes===		===Notes===
-	* Our "1X TBE" is technically 0.5X TBE but for our purposes we call it 1X TBE.	+	* Our "1X TBE" is technically 0.5X TBE (5.3g/L Tris Base, 2.75g/L Boric Acid, and 2.9g/L EDTA) but for our purposes we call it 1X TBE.
	** Correspondingly, the 10X TBE is officially 5X.		** Correspondingly, the 10X TBE is officially 5X.
	*Using TBE allows us to have such a high (150V) voltage. If you use TAE you need to use a significantly lower voltage and your run time will be longer. Despite that, TAE is advantagous in some cases, but [[User:Michael_A. Speer \| Mike]] feels that TBE is better suited for his applications. This is discussed in detail in the [[agarose gel electrophoresis \| consensus protocol]].		*Using TBE allows us to have such a high (150V) voltage. If you use TAE you need to use a significantly lower voltage and your run time will be longer. Despite that, TAE is advantagous in some cases, but [[User:Michael_A. Speer \| Mike]] feels that TBE is better suited for his applications. This is discussed in detail in the [[agarose gel electrophoresis \| consensus protocol]].

2009-06-23T17:49:48Z

←Older revision		Revision as of 17:49, 23 June 2009
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	<center>		<center>
	'''This protocol is for the typical separation and purification of DNA done in the Richard Lab.		'''This protocol is for the typical separation and purification of DNA done in the Richard Lab.
-	'''The [[agarose gel electrophoresis \| consensus electrophoresis protocol]] should be consulted if deviating from the procedure outlined here.'''</center>	+	'''The [[agarose gel electrophoresis \| consensus electrophoresis protocol]] should be consulted if using different reagents or otherwise deviating from the procedure outlined here.'''</center>

	===Procedure===		===Procedure===

2009-06-23T17:45:27Z

Notes

2009-06-23T17:45:05Z

Notes

←Older revision		Revision as of 17:45, 23 June 2009
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	! Agarose Concentration (g/100mL) !! Optimal DNA Resolution (kb)		! Agarose Concentration (g/100mL) !! Optimal DNA Resolution (kb)
	\|-=		\|-=
-	\| 0.5 \|\| 1 - 30	+	\| align="center"\|0.5 \|\| align="center"\|1 - 30
	\|-		\|-
	\| 0.7 \|\| 0.8 - 12		\| 0.7 \|\| 0.8 - 12

2009-06-23T17:44:02Z

←Older revision		Revision as of 17:44, 23 June 2009
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-	Back to [[Richard_Lab \| ~~Richard Lab~~]]	+	Back to [[Richard_Lab:protocols \| Protocols]]

	[[Image:Gel_electrophoresis.JPG\|thumb\|center\|300px]]		[[Image:Gel_electrophoresis.JPG\|thumb\|center\|300px]]

2009-06-23T17:43:28Z

Notes

←Older revision		Revision as of 17:43, 23 June 2009
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	* Use the following table to determine the amount of agarose you want to use.		* Use the following table to determine the amount of agarose you want to use.
	<center>		<center>
-	{\| border="1" cellpadding="5" cellspacing="0" align="center"	+	{\| border="1" cellpadding="5" cellspacing="0" align="center"
	\|+ '''Choosing an Agarose Concentration'''		\|+ '''Choosing an Agarose Concentration'''
	! Agarose Concentration (g/100mL) !! Optimal DNA Resolution (kb)		! Agarose Concentration (g/100mL) !! Optimal DNA Resolution (kb)