Haynes Lab:Notebook/Engineering PC-TFs/2012/12/07 - Revision history

Brady S. Laughlin: /* Summary */

2012-12-07T23:04:05Z

Summary

←Older revision		Revision as of 23:04, 7 December 2012
Line 70:		Line 70:
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	*Ran Zymoclean DNA recovery kit.		*Ran Zymoclean DNA recovery kit.
		+
		+	*Add 3 volumes of ADB Buffer to each volume of gel. (Each gel slice is assumed to be 200 mg)
		+	*Therefore, add 600 μL of ADB Buffer.
		+	*Incubate at 55°C for 10 minutes.
		+	*Load melted agarose solution into Spin Column in a collection tube.
		+	*Centrifuge at max speed for 30 seconds.
		+	*Discard flow-through.
		+	*Add 200 μL of DNA wash buffer to the spin columns and centrifuge for 30 seconds.
		+	*Place spin column in a 1.5 mL tube. Add 10μL of DNA Elution Buffer to elute the DNA.
		+	*Place tube into freezer box.
		+
		+	''There was not much DNA that came from the purification kit. A reason for this is because the bands were smaller than usual, according to Vi. Vi is running another gel digest to cut the backbone from the PSB1A3 vector. Rene said she was also running into problems with minipreping the vectors and cutting it. A follow-up with Dr. Haynes will confirm next step and possible options.
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Brady S. Laughlin: /* Summary */

2012-12-07T23:00:30Z

Summary

←Older revision		Revision as of 23:00, 7 December 2012
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-		+	*Ran Zymoclean DNA recovery kit.
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Brady S. Laughlin: /* Summary */

2012-12-07T22:59:28Z

Summary

←Older revision		Revision as of 22:59, 7 December 2012
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	PCR was set with preset settings:		PCR was set with preset settings:

-	[[Image:pcrsett.jpg\|300px~~\|thumb\|~~]]	+	[[Image:pcrsett.jpg\|300px]]

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Brady S. Laughlin: /* Summary */

2012-12-07T22:58:36Z

Summary

←Older revision		Revision as of 22:58, 7 December 2012
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	PCR was set with preset settings:		PCR was set with preset settings:
		+
		+	[[Image:pcrsett.jpg\|300px\|thumb\|]]

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Brady S. Laughlin: /* Summary */

2012-12-07T22:54:52Z

Summary

←Older revision		Revision as of 22:54, 7 December 2012
Line 10:		Line 10:


		+	*Assemblies

		+	1. hPCD - Pflex - BL01<br>
		+	2. hPCD - BL01
		+
		+	{\|border="1" cellpadding="5" cellspacing="0"
		+	\|-
		+	! scope="col" style="background:#efefef;" \| #
		+	! scope="col" style="background:#efefef;" \| Template
		+	! scope="col" style="background:#efefef;" \| Primers
		+	\|-
		+	\|1
		+	\|hPCD
		+	\|9, 2
		+	\|-
		+	\|2
		+	\|Plflex
		+	\|3, 4
		+	\|-
		+	\|3
		+	\|BL01
		+	\|5,10
		+	\|-
		+	\|4
		+	\|hPCD
		+	\|9, 7
		+	\|-
		+	\|5
		+	\|BL01
		+	\|8,10
		+	\|-
		+	\|}
		+
		+	{\|border="1" cellpadding="5" cellspacing="0"
		+	\|-
		+	! scope="col" style="background:#efefef;" \|
		+	! scope="col" style="background:#efefef;" \| μL
		+	\|-
		+	\|Template
		+	\|.5 μL
		+	\|-
		+	\|Primer 1 (10 μm)
		+	\|1.0 μL
		+	\|-
		+	\|Primer 2 (10 μm)
		+	\|1.0 μL
		+	\|-
		+	\|2*GoTaq
		+	\|12. 5
		+	\|-
		+	\|DH2O
		+	\|10.0 μL
		+	\|-
		+	\|}
		+	PCR was set with preset settings:
		+
		+	----

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Brady S. Laughlin: /* Summary */

2012-12-07T22:53:36Z

Summary

←Older revision		Revision as of 22:53, 7 December 2012
Line 7:		Line 7:
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	==Summary==		==Summary==
-	*	+	* Performed another PCR run because lid was not closed entirely yesterday. Next time in lab(Monday), confirmation gel will be run to show that PCR rxns worked.
		+
		+

Brady S. Laughlin: Autocreate 2012/12/07 Entry for Haynes_Lab:Notebook/Engineering_PC-TFs

2012-12-07T22:52:18Z

Autocreate 2012/12/07 Entry for Haynes_Lab:Notebook/Engineering_PC-TFs

New page

{|{{table}} width="800"
|-
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Engineering PC-TFs</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>      </html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|-
| colspan="2"|

==Summary==
*


|}

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