Will Notebook1 - Revision history

Wideloache: /* Wideloache 14:06, 19 December 2008 (EST) */

2008-12-19T19:27:58Z

‎Wideloache 14:06, 19 December 2008 (EST)

2008-12-19T19:06:49Z

2008-12-19T18:59:52Z

2008-12-18T21:52:42Z

2008-12-18T21:51:26Z

2008-12-18T21:47:37Z

2008-12-18T05:32:50Z

2008-12-11T21:34:30Z

2008-12-11T21:33:42Z

2008-12-11T20:21:25Z

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 Move pLux and pLuxLas into pSB1A2. Put all 4 promoters in front of GFP.
 Complete the genomic insertion. Try different growth conditions at 43C to see if I can get the final step of the procedure to work.
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 [[Will_DeLoache Notebook|Back to Main Page]]
-==~~==
+==[[User:Wideloache|Wideloache]] 14:06, 19 December 2008 (EST)==
 Today I ran the O/N digestions of pSB1A2-pLux' and pSB1A2-pLuxLas on a gel:
 [[Image:2008-12-19 13hr 21min.jpg|thumb|none|100px|1.6% gel. Lane 1: pSB1A2-pLux'(E/P). Lane 2: pSB1A2-pLuxLas (E/P).]]
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 [[Will_DeLoache Notebook|Back to Main Page]]
 ==[[User:Wideloache|Wideloache]] 16:51, 18 December 2008 (EST)==
 Done with exams finally!
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 Ec100D::pir+ +pG80ko-K091206
 '''Kanamycin:'''

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 I picked colonies from the following plates for O/N growth in the respective antibiotics + LB. I will freeze these cells down tomorrow and enter them into the database:
-Ampicillin:
+'''Ampicillin:'''
 pSB1A2-K091204
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 HB101 +pInt80-649
-Gentamicin:
+'''Gentamicin:'''
 Ec100D::pir+ +pG80ko-stufferInsert
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 HB101 +pInt80-649 +pG80ko-K091206
-Kanamycin:
+'''Kanamycin:'''
 LacIX86-pLux'
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 LacII12X86-pLuxLas
-LB:
+'''LB:'''
 Ec100D::pir+

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 [[Will_DeLoache Notebook|Back to Main Page]]
-==~~==
+==[[User:Wideloache|Wideloache]] 16:51, 18 December 2008 (EST)==
-Done with exams finally! Today I picked colonies from the following plates for O/N growth in the respective antibiotics + LB. I will freeze these cells down tomorrow and enter them into the database:
+Done with exams finally!
 Ampicillin:
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 pSB1A2-B0015
-pSB1A2-pLux-1 (The second colony didn't grow, see below)
+pSB1A2-pLux-1 (The second colony didn't grow, see above)
 pSB1A2-pLuxLas-1 (The second colony didn't grow)

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 [[Image:2008-12-11 14hr 49min.jpg|thumb|150px|none|.4% gel. All plasmids cut with EcoRI. Lane 1: Miniprep of HB101 cells on LB+Gen plate. Lane 2: pInt80-649. Lane 3: pG80ko-K091206.]]
-Expected lengths for the two plasmids were: pInt80-649 = 6335bp. pG80ko-K091206 = 4268bp. Both plamids appear to be in the miniprep sample, although other bands are present at 2000bp and possibly at 1000.
+Expected lengths for the two plasmids were: pInt80-649 = 6335bp. pG80ko-K091206 = 4268bp. Both plamids appear to be in the miniprep sample, although other bands are present at 2000bp and possibly at 1000. There is also a band that is larger than expected above pInt80-649.
 [[Image:2008-12-11 14hr 38min.jpg|thumb|200px|none|2.0% gel. Lane 1: LacIX86-pLux' (E). Lane 2: LacII12X86-pLuxLas (E). Lane 3: LacIX86-pLux'  - MP (E/P). Lane 4: LacII12X86-pLuxLas - MP (E/P)]]

← Older revision		Revision as of 21:33, 11 December 2008
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	[[Image:2008-12-11 14hr 49min.jpg\|thumb\|150px\|none\|.4% gel. All plasmids cut with EcoRI. Lane 1: Miniprep of HB101 cells on LB+Gen plate. Lane 2: pInt80-649. Lane 3: pG80ko-K091206.]]		[[Image:2008-12-11 14hr 49min.jpg\|thumb\|150px\|none\|.4% gel. All plasmids cut with EcoRI. Lane 1: Miniprep of HB101 cells on LB+Gen plate. Lane 2: pInt80-649. Lane 3: pG80ko-K091206.]]
		+	Expected lengths for the two plasmids were: pInt80-649 = 6335bp. pG80ko-K091206 = 4268bp. Both plamids appear to be in the miniprep sample, although other bands are present at 2000bp and possibly at 1000.

	[[Image:2008-12-11 14hr 38min.jpg\|thumb\|200px\|none\|2.0% gel. Lane 1: LacIX86-pLux' (E). Lane 2: LacII12X86-pLuxLas (E). Lane 3: LacIX86-pLux' - MP (E/P). Lane 4: LacII12X86-pLuxLas - MP (E/P)]]		[[Image:2008-12-11 14hr 38min.jpg\|thumb\|200px\|none\|2.0% gel. Lane 1: LacIX86-pLux' (E). Lane 2: LacII12X86-pLuxLas (E). Lane 3: LacIX86-pLux' - MP (E/P). Lane 4: LacII12X86-pLuxLas - MP (E/P)]]

← Older revision		Revision as of 20:21, 11 December 2008
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	[[Image:2008-12-11 14hr 38min.jpg\|thumb\|200px\|none\|2.0% gel. Lane 1: LacIX86-pLux' (E). Lane 2: LacII12X86-pLuxLas (E). Lane 3: LacIX86-pLux' - MP (E/P). Lane 4: LacII12X86-pLuxLas - MP (E/P)]]		[[Image:2008-12-11 14hr 38min.jpg\|thumb\|200px\|none\|2.0% gel. Lane 1: LacIX86-pLux' (E). Lane 2: LacII12X86-pLuxLas (E). Lane 3: LacIX86-pLux' - MP (E/P). Lane 4: LacII12X86-pLuxLas - MP (E/P)]]
		+
		+	The digestion of the two synthesized parts was successful. The plasmid from GeneArt and the plasmid from the miniprep appeared to be the same. Because the miniprepped plasmid had been cut with PstI as well as EcoRI, the vector had been cut in half and my two promoter parts were a little shorter. I excised these two shorter bands and ligated into pSB1A2.
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