User:Karmella Haynes/Notebook/BioBrick cloning/2012/12/11 - Revision history

Karmella Haynes: /* 12/11/12 */

2012-12-12T18:27:05Z

12/11/12

←Older revision		Revision as of 18:27, 12 December 2012
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	----		----
	'''Assemblies'''		'''Assemblies'''
-	# KAH111/C-His: 5' part/(E/P)/~~size~~ + C-His/(E/P)/3600	+	# KAH111/C-His: 5' part/(E/P)/1123 + C-His/(E/P)/3600


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	\| Ligation \|\| <font color="blue"><u>Plate results (lig : neg crtl)</u> 12/12/12</font>		\| Ligation \|\| <font color="blue"><u>Plate results (lig : neg crtl)</u> 12/12/12</font>
	\|-		\|-
-	\| 1. KAH111(E/P)/size, 16 ng + C-His(E/P)/3600, 25ng \|\| <font color="blue">~~new BioBrick #~~:1 (Pick #)</font>	+	\| 1. KAH111(E/P)/size, 16 ng + C-His(E/P)/3600, 25ng \|\| <font color="blue">KAH111/C-His 3:1 (Pick 4)</font> (very small colonies after ~18 hours)
	\|-		\|-
-	\| 2. C-His(E/P)/ 25 ng \|\| ~~ ~~	+	\| 2. C-His(E/P)/ 25 ng \|\| 3 colonies
	\|}		\|}

Karmella Haynes: /* 12/11/12 */

2012-12-12T18:25:31Z

12/11/12

←Older revision		Revision as of 18:25, 12 December 2012
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	\| bgcolor=#cfcfcf \| Reagent		\| bgcolor=#cfcfcf \| Reagent
	\| bgcolor=#cfcfcf \| Volume		\| bgcolor=#cfcfcf \| Volume
-	\| rowspan="7" \| <u>Expected:</u><br>3. pSB1A3 = ~~size~~<br>4. ~~BB2~~ = ~~size~~<br>	+	\| rowspan="7" \| <u>Expected:</u><br>3. pSB1A3 = 2155<br>4. pSB1A3 = 2155<br>
	\| rowspan="7" \| [[Image:KAH121112_gel1.png\|200px\|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]		\| rowspan="7" \| [[Image:KAH121112_gel1.png\|200px\|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
	\|-		\|-

Karmella Haynes: /* 12/11/12 */

2012-12-12T00:14:00Z

12/11/12

←Older revision		Revision as of 00:14, 12 December 2012
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	* Digests (Fermentas FD)		* Digests (Fermentas FD)
-	** Specific notes

	{\| {{table}} cellspacing="3" <!-- Digest rxn. table -->		{\| {{table}} cellspacing="3" <!-- Digest rxn. table -->
Line 47:		Line 46:
	\| Sample \|\| OD260 \|\| 260/280 \|\| ng/μL		\| Sample \|\| OD260 \|\| 260/280 \|\| ng/μL
	\|-		\|-
-	\| 1. KAH11 (E/P) \|\| ~~---~~ \|\| ~~---~~ \|\| ~~---~~	+	\| 1. KAH11 (E/P) \|\| 0.019 \|\| 1.96 \|\| 18.7
	\|-		\|-
-	\| 2. C-His (E/P) \|\| ~~---~~ \|\| ~~---~~ \|\| ~~---~~	+	\| 2. C-His (E/P) \|\| 0.047 \|\| 1.82 \|\| 46.8
	\|}		\|}

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	* pSB1A3-RFP (retransformation from 12/10/12)		* pSB1A3-RFP (retransformation from 12/10/12)
	* Note: pellets are red		* Note: pellets are red
-	* Concentrations: (1) 260 = , 260/280 = , '''ng/μL = '''; (2) 260 = , 260/280 = , '''ng/μL = ''';
	* Digest 10 μL of each with X/S for Gibson cloning (Brady and Vi)		* Digest 10 μL of each with X/S for Gibson cloning (Brady and Vi)
		+	* Note: combined mini preps afterwards; concentration = 60 ng/μL

	{\| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table -->		{\| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table -->
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	* Gel purification: combined the two slices		* Gel purification: combined the two slices
-	** Concentration: 260 = , 260/280 = , '''ng/μL = '''	+	** Concentration: 260 = 0.015, 260/280 = 2.2, '''ng/μL = 15.1'''

Karmella Haynes: /* 12/11/12 */

2012-12-11T23:38:59Z

12/11/12

←Older revision		Revision as of 23:38, 11 December 2012
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	==12/11/12==		==12/11/12==
	<!-- Precede finished items with a checkmark ✓ -->		<!-- Precede finished items with a checkmark ✓ -->
		+	* Note: colonies from retransformation of pSB1A3-RFP look red
	* Assembly: KAH111 + C-His (trouble shooting for Behzad, start from plasmids)		* Assembly: KAH111 + C-His (trouble shooting for Behzad, start from plasmids)
	* Minipreps: Rene's pSB1A3-RFP (2); cut with X/S for Brady and Vi's Gibson cloning		* Minipreps: Rene's pSB1A3-RFP (2); cut with X/S for Brady and Vi's Gibson cloning
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	'''Minipreps'''<br>		'''Minipreps'''<br>
	* pSB1A3-RFP (retransformation from 12/10/12)		* pSB1A3-RFP (retransformation from 12/10/12)
		+	* Note: pellets are red
	* Concentrations: (1) 260 = , 260/280 = , '''ng/μL = '''; (2) 260 = , 260/280 = , '''ng/μL = ''';		* Concentrations: (1) 260 = , 260/280 = , '''ng/μL = '''; (2) 260 = , 260/280 = , '''ng/μL = ''';
	* Digest 10 μL of each with X/S for Gibson cloning (Brady and Vi)		* Digest 10 μL of each with X/S for Gibson cloning (Brady and Vi)

Karmella Haynes: /* 12/11/12 */

2012-12-11T23:33:56Z

12/11/12

←Older revision		Revision as of 23:33, 11 December 2012
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	{\| {{table}} cellspacing="3" <!-- Ligation rxn table -->		{\| {{table}} cellspacing="3" <!-- Ligation rxn table -->
-	\|   \|\| 1 \|\| 2 \|\|	+	\|   \|\| 1 \|\| 2
	\|-		\|-
-	\| Insert DNA \|\| 1.0 \|\| --- \|\|	+	\| Insert DNA \|\| 1.0 \|\| ---
	\|-		\|-
-	\| Vector DNA \|\| 0.5 \|\| 0.5 \|\|	+	\| Vector DNA \|\| 0.5 \|\| 0.5
	\|-		\|-
-	\| 2x lgn buf (Roche) \|\| 5.0 \|\| 5.0 \|\|	+	\| 2x lgn buf (Roche) \|\| 5.0 \|\| 5.0
	\|-		\|-
-	\| T4 ligase (NEB) \|\| 1.0 \|\| 1.0 \|\|	+	\| T4 ligase (NEB) \|\| 1.0 \|\| 1.0
	\|-		\|-
-	\| dH<sub>2</sub>O \|\| 2.5 \|\| 3.5 \|\|	+	\| dH<sub>2</sub>O \|\| 2.5 \|\| 3.5
	\|-		\|-
-	\|   \|\| 10 μL \|\| 10 μL \|\|	+	\|   \|\| 10 μL \|\| 10 μL
	\|}		\|}

Karmella Haynes: /* 12/11/12 */

2012-12-11T23:33:31Z

12/11/12

←Older revision		Revision as of 23:33, 11 December 2012
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-	* Measure conc.'s	+	* Measure conc.'s of DNA purified from gel (Zymo kit)
	{\| {{table}} cellspacing="3" <!-- [DNA] table -->		{\| {{table}} cellspacing="3" <!-- [DNA] table -->
	\|- bgcolor=#cfcfcf		\|- bgcolor=#cfcfcf

Karmella Haynes: /* 12/11/12 */

2012-12-11T23:32:36Z

12/11/12

←Older revision		Revision as of 23:32, 11 December 2012
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	\| bgcolor=#cfcfcf \| Reagent		\| bgcolor=#cfcfcf \| Reagent
	\| bgcolor=#cfcfcf \| Volume		\| bgcolor=#cfcfcf \| Volume
-	\| rowspan="7" \| [[Image:~~KAH121112_gel1~~.png\|200px\|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]	+	\| rowspan="7" \| [[Image:KAH121112_gel2.png\|200px\|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
	\|-		\|-
	\| DNA (plasmid) \|\| 15.0 μL		\| DNA (plasmid) \|\| 15.0 μL
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	\| bgcolor=#cfcfcf \| Volume		\| bgcolor=#cfcfcf \| Volume
	\| rowspan="7" \| <u>Expected:</u><br>3. pSB1A3 = size<br>4. BB2 = size<br>		\| rowspan="7" \| <u>Expected:</u><br>3. pSB1A3 = size<br>4. BB2 = size<br>
-	\| rowspan="7" \| [[Image:~~KAH121112_gel2~~.png\|200px\|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]	+	\| rowspan="7" \| [[Image:KAH121112_gel1.png\|200px\|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
	\|-		\|-
	\| DNA(plasmid) \|\| 10.0 μL		\| DNA(plasmid) \|\| 10.0 μL
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	\|   \|\| 15 μL --> 37°C/ ~30 min.		\|   \|\| 15 μL --> 37°C/ ~30 min.
	\|}		\|}
		+
		+
		+	* Gel purification: combined the two slices
		+	** Concentration: 260 = , 260/280 = , '''ng/μL = '''

Karmella Haynes: /* 12/11/12 */

2012-12-11T23:30:46Z

12/11/12

←Older revision		Revision as of 23:30, 11 December 2012
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	\| bgcolor=#cfcfcf \| Reagent		\| bgcolor=#cfcfcf \| Reagent
	\| bgcolor=#cfcfcf \| Volume		\| bgcolor=#cfcfcf \| Volume
-	\| rowspan="7" \| [[Image:KAH121112_gel1.png\|~~150px~~\|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]	+	\| rowspan="7" \| [[Image:KAH121112_gel1.png\|200px\|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
	\|-		\|-
	\| DNA (plasmid) \|\| 15.0 μL		\| DNA (plasmid) \|\| 15.0 μL
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	\| bgcolor=#cfcfcf \| Volume		\| bgcolor=#cfcfcf \| Volume
	\| rowspan="7" \| <u>Expected:</u><br>3. pSB1A3 = size<br>4. BB2 = size<br>		\| rowspan="7" \| <u>Expected:</u><br>3. pSB1A3 = size<br>4. BB2 = size<br>
-	\| rowspan="7" \| [[Image:KAH121112_gel2.png\|~~150px~~\|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]	+	\| rowspan="7" \| [[Image:KAH121112_gel2.png\|200px\|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
	\|-		\|-
	\| DNA(plasmid) \|\| 10.0 μL		\| DNA(plasmid) \|\| 10.0 μL

Karmella Haynes: /* 12/11/12 */

2012-12-11T23:30:22Z

12/11/12

←Older revision		Revision as of 23:30, 11 December 2012
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	\| bgcolor=#cfcfcf \| Reagent		\| bgcolor=#cfcfcf \| Reagent
	\| bgcolor=#cfcfcf \| Volume		\| bgcolor=#cfcfcf \| Volume
-	\| rowspan="7" \| ~~<!--~~ [[Image:KAH121112_gel1.png\|~~250px~~\|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] ~~-->~~	+	\| rowspan="7" \| [[Image:KAH121112_gel1.png\|150px\|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
	\|-		\|-
	\| DNA (plasmid) \|\| 15.0 μL		\| DNA (plasmid) \|\| 15.0 μL
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	\| bgcolor=#cfcfcf \| Volume		\| bgcolor=#cfcfcf \| Volume
	\| rowspan="7" \| <u>Expected:</u><br>3. pSB1A3 = size<br>4. BB2 = size<br>		\| rowspan="7" \| <u>Expected:</u><br>3. pSB1A3 = size<br>4. BB2 = size<br>
-	\| rowspan="7" \| [[Image:KAH121112_gel2.png\|~~400px~~\|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]	+	\| rowspan="7" \| [[Image:KAH121112_gel2.png\|150px\|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
	\|-		\|-
	\| DNA(plasmid) \|\| 10.0 μL		\| DNA(plasmid) \|\| 10.0 μL

Karmella Haynes: /* 12/11/12 */

2012-12-11T23:29:12Z

12/11/12

←Older revision		Revision as of 23:29, 11 December 2012
Line 12:		Line 12:


-	~~----~~
-	~~'''Minipreps'''<br>~~
-	* pSB1A3-RFP (retransformation from 12/10/12)
-	* Concentrations: (1) 260 = , 260/280 = , '''ng/μL = '''; (2) 260 = , 260/280 = , '''ng/μL = ''';
-	* Digest 10 μL of each with X/S for Gibson cloning (Brady and Vi)
-
-	~~{\| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table -->~~
-	~~\|- valign="top"~~
-	~~\| bgcolor=#cfcfcf \| Reagent~~
-	~~\| bgcolor=#cfcfcf \| Volume~~
-	~~\| rowspan="7" \| <u>Expected:</u><br>3. pSB1A3 = size<br>4. BB2 = size<br>~~
-	~~\| rowspan="7" \| [[Image:KAH121112_gel1.png\|400px\|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]~~
-	\|-
-	~~\| DNA(plasmid) \|\| 10.0 μL~~
-	\|-
-	~~\| 10X buffer \|\| 1.5~~
-	\|-
-	~~\| EcoRI \|\| 1.0~~
-	\|-
-	~~\| PstI \|\| 1.0~~
-	\|-
-	~~\| dH<sub>2</sub>O \|\| 9.5~~
-	\|-
-	~~\|   \|\| 15 μL --> 37°C/ ~15 min.~~
-	\|}

	----		----
	'''Assemblies'''		'''Assemblies'''
-	# ~~BioBrick name: 5' part~~/~~(a/b)/size + 3' part/(c/d)/size~~	+	# KAH111/C-His: 5' part/(E/P)/size + C-His/(E/P)/3600
-	~~# BioBrick name~~: 5' part/(a/b)/size + ~~3' part~~/(c/d)/~~size~~	+


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	\| bgcolor=#cfcfcf \| Reagent		\| bgcolor=#cfcfcf \| Reagent
	\| bgcolor=#cfcfcf \| Volume		\| bgcolor=#cfcfcf \| Volume
-	\| rowspan="7" \| <!-- [[Image:~~GelImage~~.~~jpg~~\|~~270px~~\|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] -->	+	\| rowspan="7" \| <!-- [[Image:KAH121112_gel1.png\|250px\|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] -->
	\|-		\|-
-	\| DNA (plasmid) \|\| ~~up to 25~~ μL	+	\| DNA (plasmid) \|\| 15.0 μL
	\|-		\|-
	\| 10x buffer \|\| 3.0		\| 10x buffer \|\| 3.0
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	\| enzyme 2 \|\| 1.0		\| enzyme 2 \|\| 1.0
	\|-		\|-
-	\| dH<sub>2</sub>O \|\| ~~---~~	+	\| dH<sub>2</sub>O \|\| 10.0
	\|-		\|-
	\|   \|\| 30 μL --> 37°C/ ~30 min.		\|   \|\| 30 μL --> 37°C/ ~30 min.
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	\| Sample \|\| OD260 \|\| 260/280 \|\| ng/μL		\| Sample \|\| OD260 \|\| 260/280 \|\| ng/μL
	\|-		\|-
-	\| 1. ~~Digested part~~ (a/b) \|\| --- \|\| --- \|\| ---	+	\| 1. KAH11 (E/P) \|\| --- \|\| --- \|\| ---
	\|-		\|-
-	\| 2. ~~Digested part~~ (c/d) \|\| --- \|\| --- \|\| ---	+	\| 2. C-His (E/P) \|\| --- \|\| --- \|\| ---
-	\|}	+
-		+
-		+
-	* Dephosphorylation (Roche)	+
-	~~{\| {{table}} cellspacing="3" <!-- Dephos table -->~~	+
-	\|-	+
-	~~\| bgcolor=#cfcfcf \| Reagent~~	+
-	~~\| bgcolor=#cfcfcf \| Volume~~	+
-	\|-	+
-	~~\| DNA (clean digest) \|\| up to 17 μL (500 ng)~~	+
-	\|-	+
-	~~\| 10x buffer d.p. \|\| 2.0~~	+
-	\|-	+
-	~~\| phosphatase \|\| 1.0~~	+
-	\|-	+
-	~~\| dH<sub>2</sub>O \|\| ---~~	+
-	\|-	+
-	~~\|   \|\| 20 μL --> 37°C/ 10 min.; 75°C/ 2 min.; [final] = 25 ng/μL~~	+
	\|}		\|}

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	{\| {{table}} cellspacing="3" <!-- Ligations table -->		{\| {{table}} cellspacing="3" <!-- Ligations table -->
	\|- bgcolor=#cfcfcf		\|- bgcolor=#cfcfcf
-	\| Ligation \|\| <font color="blue"><u>Plate results (lig : neg crtl)</u> mm/dd/yy</font>	+	\| Ligation \|\| <font color="blue"><u>Plate results (lig : neg crtl)</u> 12/12/12</font>
	\|-		\|-
-	\| 1. ~~insert~~(a/b)/size, ## ng + ~~vector~~(c/d)/~~size~~, ~~## ng~~ \|\| <font color="blue">new BioBrick #:1 (Pick #)</font>	+	\| 1. KAH111(E/P)/size, 16 ng + C-His(E/P)/3600, 25ng \|\| <font color="blue">new BioBrick #:1 (Pick #)</font>
	\|-		\|-
-	\| 2. ~~vector~~(c/d)/ ## ng \|\|	+	\| 2. C-His(E/P)/ 25 ng \|\|
	\|}		\|}

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	\|   \|\| 1 \|\| 2 \|\|		\|   \|\| 1 \|\| 2 \|\|
	\|-		\|-
-	\| Insert DNA \|\| ~~###~~ \|\| --- \|\|	+	\| Insert DNA \|\| 1.0 \|\| --- \|\|
	\|-		\|-
-	\| Vector DNA \|\| ~~###~~ \|\| ~~###~~ \|\|	+	\| Vector DNA \|\| 0.5 \|\| 0.5 \|\|
	\|-		\|-
-	\| 2x lgn buf (Roche) \|\| ~~###~~ \|\| ~~###~~ \|\|	+	\| 2x lgn buf (Roche) \|\| 5.0 \|\| 5.0 \|\|
	\|-		\|-
	\| T4 ligase (NEB) \|\| 1.0 \|\| 1.0 \|\|		\| T4 ligase (NEB) \|\| 1.0 \|\| 1.0 \|\|
	\|-		\|-
-	\| dH<sub>2</sub>O \|\| ~~###~~ \|\| ~~###~~ \|\|	+	\| dH<sub>2</sub>O \|\| 2.5 \|\| 3.5 \|\|
	\|-		\|-
-	\|   \|\| # μL \|\| # μL \|\|	+	\|   \|\| 10 μL \|\| 10 μL \|\|
	\|}		\|}
		+
		+

	----		----
-	'''~~Oligo annealing~~'''	+	'''Minipreps'''<br>
-	~~# New BB 1~~	+	* pSB1A3-RFP (retransformation from 12/10/12)
-	~~# New BB~~ 2	+	* Concentrations: (1) 260 = , 260/280 = , '''ng/μL = '''; (2) 260 = , 260/280 = , '''ng/μL = ''';
		+	* Digest 10 μL of each with X/S for Gibson cloning (Brady and Vi)

-	{\| ~~class="wikitable"~~ border="0" cellspacing="3" <!-- ~~Oligo annealing~~ rxn table -->	+	{\| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table -->
-	\| DNA (~~oligos, 100 μM~~) \|\| ~~up to 18~~ μL (3 ~~μL ea~~.)	+	\|- valign="top"
		+	\| bgcolor=#cfcfcf \| Reagent
		+	\| bgcolor=#cfcfcf \| Volume
		+	\| rowspan="7" \| <u>Expected:</u><br>3. pSB1A3 = size<br>4. BB2 = size<br>
		+	\| rowspan="7" \| [[Image:KAH121112_gel2.png\|400px\|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
		+	\|-
		+	\| DNA(plasmid) \|\| 10.0 μL
		+	\|-
		+	\| 10X buffer \|\| 3.0
		+	\|-
		+	\| XbaI \|\| 1.0
	\|-		\|-
-	\| ~~10x annealing buffer~~ \|\| 2.0	+	\| SpeI \|\| 1.0
	\|-		\|-
-	\| dH<sub>2</sub>O \|\| ~~---~~	+	\| dH<sub>2</sub>O \|\| 15.0
	\|-		\|-
-	\|   \|\| 20 μL --> ~~100°C (water bath)~~/ 5 min.~~; Cool to R.T. overnight~~	+	\|   \|\| 15 μL --> 37°C/ ~30 min.
	\|}		\|}

←Older revision		Revision as of 18:25, 12 December 2012
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	\| bgcolor=#cfcfcf \| Reagent		\| bgcolor=#cfcfcf \| Reagent
	\| bgcolor=#cfcfcf \| Volume		\| bgcolor=#cfcfcf \| Volume
-	\| rowspan="7" \| <u>Expected:</u><br>3. pSB1A3 = ~~size~~<br>4. ~~BB2~~ = ~~size~~<br>	+	\| rowspan="7" \| <u>Expected:</u><br>3. pSB1A3 = 2155<br>4. pSB1A3 = 2155<br>
	\| rowspan="7" \| [[Image:KAH121112_gel1.png\|200px\|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]		\| rowspan="7" \| [[Image:KAH121112_gel1.png\|200px\|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
	\|-		\|-

←Older revision		Revision as of 23:32, 11 December 2012
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	\| bgcolor=#cfcfcf \| Reagent		\| bgcolor=#cfcfcf \| Reagent
	\| bgcolor=#cfcfcf \| Volume		\| bgcolor=#cfcfcf \| Volume
-	\| rowspan="7" \| [[Image:~~KAH121112_gel1~~.png\|200px\|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]	+	\| rowspan="7" \| [[Image:KAH121112_gel2.png\|200px\|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
	\|-		\|-
	\| DNA (plasmid) \|\| 15.0 μL		\| DNA (plasmid) \|\| 15.0 μL
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	\| bgcolor=#cfcfcf \| Volume		\| bgcolor=#cfcfcf \| Volume
	\| rowspan="7" \| <u>Expected:</u><br>3. pSB1A3 = size<br>4. BB2 = size<br>		\| rowspan="7" \| <u>Expected:</u><br>3. pSB1A3 = size<br>4. BB2 = size<br>
-	\| rowspan="7" \| [[Image:~~KAH121112_gel2~~.png\|200px\|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]	+	\| rowspan="7" \| [[Image:KAH121112_gel1.png\|200px\|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
	\|-		\|-
	\| DNA(plasmid) \|\| 10.0 μL		\| DNA(plasmid) \|\| 10.0 μL
Line 105:		Line 105:
	\|   \|\| 15 μL --> 37°C/ ~30 min.		\|   \|\| 15 μL --> 37°C/ ~30 min.
	\|}		\|}
		+
		+
		+	* Gel purification: combined the two slices
		+	** Concentration: 260 = , 260/280 = , '''ng/μL = '''

←Older revision		Revision as of 23:30, 11 December 2012
Line 25:		Line 25:
	\| bgcolor=#cfcfcf \| Reagent		\| bgcolor=#cfcfcf \| Reagent
	\| bgcolor=#cfcfcf \| Volume		\| bgcolor=#cfcfcf \| Volume
-	\| rowspan="7" \| [[Image:KAH121112_gel1.png\|~~150px~~\|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]	+	\| rowspan="7" \| [[Image:KAH121112_gel1.png\|200px\|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
	\|-		\|-
	\| DNA (plasmid) \|\| 15.0 μL		\| DNA (plasmid) \|\| 15.0 μL
Line 91:		Line 91:
	\| bgcolor=#cfcfcf \| Volume		\| bgcolor=#cfcfcf \| Volume
	\| rowspan="7" \| <u>Expected:</u><br>3. pSB1A3 = size<br>4. BB2 = size<br>		\| rowspan="7" \| <u>Expected:</u><br>3. pSB1A3 = size<br>4. BB2 = size<br>
-	\| rowspan="7" \| [[Image:KAH121112_gel2.png\|~~150px~~\|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]	+	\| rowspan="7" \| [[Image:KAH121112_gel2.png\|200px\|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
	\|-		\|-
	\| DNA(plasmid) \|\| 10.0 μL		\| DNA(plasmid) \|\| 10.0 μL

←Older revision		Revision as of 23:30, 11 December 2012
Line 25:		Line 25:
	\| bgcolor=#cfcfcf \| Reagent		\| bgcolor=#cfcfcf \| Reagent
	\| bgcolor=#cfcfcf \| Volume		\| bgcolor=#cfcfcf \| Volume
-	\| rowspan="7" \| ~~<!--~~ [[Image:KAH121112_gel1.png\|~~250px~~\|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] ~~-->~~	+	\| rowspan="7" \| [[Image:KAH121112_gel1.png\|150px\|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
	\|-		\|-
	\| DNA (plasmid) \|\| 15.0 μL		\| DNA (plasmid) \|\| 15.0 μL
Line 91:		Line 91:
	\| bgcolor=#cfcfcf \| Volume		\| bgcolor=#cfcfcf \| Volume
	\| rowspan="7" \| <u>Expected:</u><br>3. pSB1A3 = size<br>4. BB2 = size<br>		\| rowspan="7" \| <u>Expected:</u><br>3. pSB1A3 = size<br>4. BB2 = size<br>
-	\| rowspan="7" \| [[Image:KAH121112_gel2.png\|~~400px~~\|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]	+	\| rowspan="7" \| [[Image:KAH121112_gel2.png\|150px\|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
	\|-		\|-
	\| DNA(plasmid) \|\| 10.0 μL		\| DNA(plasmid) \|\| 10.0 μL