Knight:DNA Spots - Revision history

Samantha L. Burke: /* Making Spots */

Samantha L. Burke — Wed, 26 Sep 2007 13:29:41 GMT

Making Spots

←Older revision		Revision as of 13:29, 26 September 2007
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	===Making Spots===		===Making Spots===
	[[Image:SpotGrid.jpg\|thumb\|left\|Spotted Grid]]		[[Image:SpotGrid.jpg\|thumb\|left\|Spotted Grid]]
-	*Mix 1 part 1% Cresol Red with 4 parts DNA that is at least 100 μL~~/ng~~	+	*Mix 1 part 1% Cresol Red with 4 parts DNA that is at least 100 ng/μL
	**''The exact amounts depend on how many 2 μL spots you plan to make''		**''The exact amounts depend on how many 2 μL spots you plan to make''
	**''Using the excess of Cresol Red is helpful for when you transform, since you can visibly tell which cells have had DNA added and which have not''		**''Using the excess of Cresol Red is helpful for when you transform, since you can visibly tell which cells have had DNA added and which have not''

Samantha L. Burke — Thu, 20 Sep 2007 20:16:10 GMT

←Older revision		Revision as of 20:16, 20 September 2007
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	*Spread cells on previously made LB plates with proper antibiotic		*Spread cells on previously made LB plates with proper antibiotic
	*Grow overnight at 37 °C		*Grow overnight at 37 °C
		+
		+	[[Category:Protocol]] [[Category:Escherichia coli]]

Samantha L. Burke — Sun, 02 Sep 2007 22:14:15 GMT

Using spots to transform E. coli

←Older revision		Revision as of 22:14, 2 September 2007
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	===Using spots to transform E. coli===		===Using spots to transform E. coli===
		+	[[Image:CuttingMat&Punch.jpg\|thumb\|left\|Olfa cutting mat and Uni-Core Punch]]
		+	[[Image:Spot.jpg\|thumb\|left\|Punched Spot]]
	*Cut out spot from surrounding paper using the Uni-Core Punch on the Olfa cutting mat		*Cut out spot from surrounding paper using the Uni-Core Punch on the Olfa cutting mat
	*Soak spot in 20 μL TE for 15 minutes.		*Soak spot in 20 μL TE for 15 minutes.

Samantha L. Burke — Sun, 02 Sep 2007 22:11:25 GMT

Making Spots

←Older revision		Revision as of 22:11, 2 September 2007
Line 12:		Line 12:

	===Making Spots===		===Making Spots===
		+	[[Image:SpotGrid.jpg\|thumb\|left\|Spotted Grid]]
	*Mix 1 part 1% Cresol Red with 4 parts DNA that is at least 100 μL/ng		*Mix 1 part 1% Cresol Red with 4 parts DNA that is at least 100 μL/ng
	**''The exact amounts depend on how many 2 μL spots you plan to make''		**''The exact amounts depend on how many 2 μL spots you plan to make''

Samantha L. Burke — Thu, 30 Aug 2007 18:45:29 GMT

Using spots to transform E. coli

←Older revision		Revision as of 18:45, 30 August 2007
Line 22:		Line 22:
	===Using spots to transform E. coli===		===Using spots to transform E. coli===
	*Cut out spot from surrounding paper using the Uni-Core Punch on the Olfa cutting mat		*Cut out spot from surrounding paper using the Uni-Core Punch on the Olfa cutting mat
-	*Soak spot in 20 μL ~~10:1~~ TE for 15 minutes.	+	*Soak spot in 20 μL TE for 15 minutes.
	*Thaw competent cells on ice while spots soak		*Thaw competent cells on ice while spots soak
	**''I used [[TOP10 chemically competent cells]]''		**''I used [[TOP10 chemically competent cells]]''

Samantha L. Burke — Thu, 30 Aug 2007 18:45:09 GMT

Materials

←Older revision		Revision as of 18:45, 30 August 2007
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	*For using spots to transform:		*For using spots to transform:
	**Harris Uni-Core Punch, 2mm and Olfa Cutting Mat		**Harris Uni-Core Punch, 2mm and Olfa Cutting Mat
-	**[TE buffer\|TE]	+	**[[TE buffer\|TE]]
	**Competent Cells		**Competent Cells

Samantha L. Burke — Thu, 30 Aug 2007 18:44:39 GMT

Materials

←Older revision		Revision as of 18:44, 30 August 2007
Line 6:		Line 6:
	*For using spots to transform:		*For using spots to transform:
	**Harris Uni-Core Punch, 2mm and Olfa Cutting Mat		**Harris Uni-Core Punch, 2mm and Olfa Cutting Mat
-	**TE	+	**[TE buffer\|TE]
	**Competent Cells		**Competent Cells

Samantha L. Burke — Thu, 30 Aug 2007 17:55:22 GMT

Materials

←Older revision		Revision as of 17:55, 30 August 2007
Line 3:		Line 3:
	**1% Cresol Red		**1% Cresol Red
	**DNA (100 ng/μL)		**DNA (100 ng/μL)
-	**[http://www.crane.com/prdSell.aspx?Name=BT886S_ThesisPapers Crane & Co.] 100% cotton, acid free thesis paper	+	**[http://www.crane.com/prdSell.aspx?Name=BT886S_ThesisPapers Crane & Co. 100% cotton, acid free thesis paper]
	*For using spots to transform:		*For using spots to transform:
	**Harris Uni-Core Punch, 2mm and Olfa Cutting Mat		**Harris Uni-Core Punch, 2mm and Olfa Cutting Mat

Samantha L. Burke — Thu, 30 Aug 2007 17:54:59 GMT

Materials

←Older revision		Revision as of 17:54, 30 August 2007
Line 3:		Line 3:
	**1% Cresol Red		**1% Cresol Red
	**DNA (100 ng/μL)		**DNA (100 ng/μL)
-	**100% cotton, acid free thesis paper	+	**[http://www.crane.com/prdSell.aspx?Name=BT886S_ThesisPapers Crane & Co.] 100% cotton, acid free thesis paper
	*For using spots to transform:		*For using spots to transform:
	**Harris Uni-Core Punch, 2mm and Olfa Cutting Mat		**Harris Uni-Core Punch, 2mm and Olfa Cutting Mat

Samantha L. Burke — Mon, 27 Aug 2007 15:35:27 GMT

Making Spots

←Older revision		Revision as of 15:35, 27 August 2007
Line 12:		Line 12:

	===Making Spots===		===Making Spots===
-	*Mix 1 part 1% Cresol Red with 4 parts ~~10-~~100 ~~ng/~~μL ~~DNA~~	+	*Mix 1 part 1% Cresol Red with 4 parts DNA that is at least 100 μL/ng
	**''The exact amounts depend on how many 2 μL spots you plan to make''		**''The exact amounts depend on how many 2 μL spots you plan to make''
	**''Using the excess of Cresol Red is helpful for when you transform, since you can visibly tell which cells have had DNA added and which have not''		**''Using the excess of Cresol Red is helpful for when you transform, since you can visibly tell which cells have had DNA added and which have not''