Team:USC/Notebook

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Lab Notebook « E. musici

Lab Notebook

5/21/12 week
-Transformed casBCED12 with Amp resistance, casBCED12 with Cm resistance, pSBIC3_RFP, pSBIK3_RFP, and pSBIA3_RFP
-successful results: pSBIC3_RFP and pSBIK3_RFP.
-inoculated pSBIC3_RFP and pSBIK3_RFP
-miniprepped pSBIK3_RFP, and pSBIT3_RFP
-all very low concentration
-Miniprepped pSBIC3_RFP, pSBIA3_RFP

-concentration (got 98 and 120 ug/ml, respectively)

-Digested pSBIC3 from pSBIC3_RFP, pSBIK3 from pSBIK3_RFP
-Transformed pSBIT3_RFP, pSBIA3_RFP, casBCDE12 on Amp
-Digested casABCDE123, tetO-GFP
-Ligated casA,3 with tetO-GFP, with pSBIA3 or pSBIC3
-Ligated casBCDE12 with pSBIA3 or PSBIC3

5/28/12 week
-Inoculated (2x) 5mL psB1A3_RFP and psB1T3_RFP.
-Transformed psB1C3_RFP, tetO_GFP, and psB3T5_RFP.
-Miniprepped psB1C3_RFP and psB1K3_RFP
-Transformed psB1C3-RFP, psB3T5-RFP, tetO-GFP
-PCRed fliE, fliM, flhD, flgA, fliD, flhB, fliA, fliE,fliF, fliG, fliH, flgJ, flgK, fliC, cheA, cheB, cheC, cheY, cheZ, luxS, lsrR, lsrK
-Gel extracted and digested the genes with EP or EX
-Miniprepped pSB3T5_RFP (3) and tetO-GFP
-Digested tetO-GFP 2

6/4/12 week
-PCRed genes using BL21 and HT115
-Gel extracted and digested
-Ligated the genes to corresponding psb1C3_RFP vectors
-Transformed 1-9F, 1-14H, 1-18H, 3-2H, 3-2J, 3-8D, 4-12A
-successful: 4-12, 3-8D, 3-2H, 1-9F, 3-2J. (very low efficiency)
-Transformed cheB, cheC, cheY, cheZ, luxS, lsrK, lsrR, flaU, flbA, flhB, flhD, flgJ, flgK, fliA, fliC, fliD, fliF, fliG, fliH, fliM
-Inoculated 4-12A, 3-8D, 3-2H, 1-9F, 3-2J
-Inoculated HT115 (w/ 5uL tet) and HB101 (w/o antibiotics)
-PCRed fliE, fliM, flhD, flgA, fliD, flhB, fliA, fliE,fliF, fliG, fliH, flgJ, flgK, fliC, cheA, cheB, cheC, cheY, cheZ, luxS, lsrR, lsrK

6/11/12 week
-Digested fliM, flhD, flgA, fliD, flhB, fliA, fliE,fliF, fliG, fliH, flgJ, flgK, fliC, cheA, cheB, cheC, cheY, cheZ, luxS, lsrR, lsrK
-Gel extracted fliM, flhD, flgA, fliD, flhB, fliA, fliE,fliF, fliG, fliH, flgJ, flgK, fliC, cheA, cheB, cheC, cheY, cheZ, luxS, lsrR, lsrK
-PCRed fliC with DH5alpha, BL21, HT115
-Ligated cm1, cm2, rfp1, rfp2
-Gel purified cm1, cm2, rfp1, rfp2
-Transformed fliA, cheZ, cheY, fliH, fliG, flgJ, flihB, flgK, fliD,fliM, 1-9F, 1-14H, 1-18H, 3-2H, 3-2J, 3-8D, 4-12A
-successful: 1-9F, 1-14H, 1-18H, 3-2H, 3-2J, 3-8D, 4-12A
-Digested cm vector with EP
-Gel extracted cm vectors
-Inoculated 3-8D, 1-9F, 4-12A
-PCRed chA, cheB, cheC, cheY, cheZ
-Transformed ligation products: fliG, flgJ, cheY, cheZ, fliD, flhB, fliM, fliH, flgK
-Ligated fliM, fliD, flhB, flgK, fliA, cheY, cheZ, fliG, flgJ, rfp1, rfp2 to Cm vector, fliH
-Miniprepped the innoculations (3-8D, 1-9F, 4-12A)
-low concentrations
-Transformed fliH, fliD, flhB, flgK, fliA, cheY, cheZ, fliG, flgJ, rfp1, rfp2, cm vector
-Miniprepped 1-9F(1), 1-9F(2), 3-8D(1), 3-8D(2), 4-12A(1), 4-12A(2)
-low concentrations

6/18/12 week
-PCRed lsrR(BL21), lsrR(HT115), lrsK(BL21), lsrK(HT115)
-Inoculated 1-9F, 3-8D, 4-12A
-Miniprepped 1-9F, 3-8D, 4-12A
-low concentrations
-Transformed J23100, RBS B0034, luxS, GFP E0040
-successful
-Inoculated J23100, RBS B0034, luxS, GFP E0040
-PCRed cheA, cheB, cheY, cheZ using HTB genomic DNA
-PCRed fliC(1), motA(2), motB(3) using DH5alpha
-Miniprepped 1-9F, 3-8D, 4-12A
-acceptable concentrations
-Digested cheA, cheB, cheY, cheZ and gel purified
-Ligated cheA, cheB, cheY, cheZ
-Transformed cheA, cheB, cheY, cheZ
-successful
-Digested psB1C3 with X and X/S
-Digested fliC, motA, motB
-Miniprepped J23100, GFP E0040, luxS, RBS Boo34
-low concentrations

6/25/12 week
-Transformed motA, motB, and control
-Inoculated T7 promoter and cheY
-Colony PCRed 20 colonies from motA, motB
-Inoculated motA, motB, cheA, cheB, cheY, cheZ
-Miniprepped cheZ, cheY, cheA, cheB, motA, motB
-acceptable concentrations
-Glycerol stock: motA 6-14, cheZ, cheY, cheA, cheB, motA, motB
-Transformed cheZ, pLaeI+RBS+cheZ, RBS+cheZ+dbl terminator, cheZ w/ H13 tag+dbl terminator, cheZ w/ H13 tag, strong promoter w/RBS
-Inoculated cheZ, pLaeI+RBS+cheZ, RBS+cheZ+dbl terminator, cheZ w/ H13 tag+dbl terminator, cheZ w/ H13 tag, strong promoter w/RBS
-PCR miniprep of cheA(7,16), cheB(14), cheZ(14), motA(15)

7/2/12 week
-Miniprepped cheZ w/ histag, cheZ, cheZ w/ RBS + dbl termination, pLacI w/ RBS + cheZ, cheZ w/ Histag + dbl termination, strong promoter w/ RBS (cm)
-acceptable concentrations
-Inoculated cheZ w/ histag (amp), cheZ (amp), cheZ w/ RBS, double termination (amp), pLacI w/ RBS, cheZ (amp), cheZ w/ histag, double termination (amp), strong promoter w/ RBS (cm)
-Miniprepped: psB1C3RFP (5 times), cheZ, strong promoter+RBS+cm, RBS+cheZ+dbl term site, pLacI+RBS+cheZ, cheZ+Hist+dbl, cheZ+Histag
-acceptable concentrations

7/9/12 week
-Digested and gel extracted fliC and GFP
-Ligated fliC
-Transformed fliC-GFP
-Received and plated strains: BW30044, BW30045, K-12 wildtype strain, strain B wildtype, JW1871-1, JW1879-2, JW2662-1, MG1655 (motile), W3110

7/16/12 week
-Made competent cells MG1655
-digested GFP vector for control using XbaI
-Colony PCRed the fliC GFP
-primer dimers. unsuccessful-

7/23/12 week
-PCRed fliC w/ RBS
-Transformed GFP-DH5a, GFP-MG1655
-Inoculated w/ 50uL transformation product
-Plated GFP-DH5a, GFP-MG1655
-PCRed fliC w/ RBS
-Miniprepped GFP-DH5a (x2), GFP-MG1655 (x2)

-acceptable concentrations

-Digested fliC and GFP w/ XbaI and gel purified
-Ligated fliC-GFP and GFP w/ H2O (control)
-Transformed 1hour ligation product
-unsuccessul
-Made control and cheY mutagenesis solutions
-Transformed fliC-GFP and GFP w/ H2O control into DH5a cells
-successful: GFP + H2O control and fliC-GFP
-Quickchange mutagenesis
-Colony PCR fliC-GFP

7/30/12 week
-Miniprepped fliC-GFP
-acceptable concentrations
-Digested miniprepped products
-Quickchange PCRed fliC-GFP
-Digested quickchange PCR products with DPN1
-Digested fliC minipreps with Spe1
-successful
-Transformed 43, 44, quickchange digests

8/6/12 week
-Digested cheY
-Quickchange PCRed fliC and cheY with control
-Digested fliC-GFP, cheY, control
-Plated fliC, cheY, and pUC18
-Inoculated cheY mut colonies
-Miniprepped cheY mut
-Glycerol stocks of cheY mut
-PCRed cheY mut
-Digested cheY mut

8/13/12 week
-PCRed fliA, fliD, fliE, fliF, fliG, fliH, fliM, flhB, flhC, flhD, flgA, flgB, flgJ, flgK, fliC-GFP, lsrR, lsrK
-Digested and gel extracted the flagella genes
-Transformed fliA, fliD,fliF, fliM, flhB, flgB, flgJ, flhD, lsrR, lsrK
-Digested fliC-GFP with DpnI, BBa_K318512, BBa_K338001, BBa_K216005
-Transformed pSB1C3 and fliC-GFP mut
-Inoculated pSB1C3-RFP, BBa_K318512, BBa_K338001, BBa_K216005
-Miniprepped pSB1C3 and BBa_K318512, BBa_K338001, BBa_K216005
-acceptable concentrations
-Digested pSB1C3 fliF, fliD, flhB, flhD, flgJ, fliM, fliA
-Ligated fliF, fliD, flhB, flhD, flgJ, fliM, fliA with pSB1C3

8/20/12 week
-Colony PCRed fliA, fliM, fliD, fliF, flgJ, flhB, flhD
-Imaged MG1655 in various conditions: pH 5, 6, 8, 9; NaCl 50mM, 100mM, 200mM, 400mM, 500mM; temperature 15C, 20C, 42C; glucose 10mM, 20mM, 50mM
-Miniprepped fliC-GFP mut, cheY mut
-acceptable concentrations
-Glycerol stocks of fliC-GFP, cheY
-Colony PCRed flgJ, flhD, fliA, flhB, cheY
-Inoculated successful colonies
-Colony PCRed fliM, fliF, fliD
-Glycerol stocks of fliA, flhB, flgJ, flhD
-Imaging of MG1655
-Miniprepped fliA, flhB, flgJ, flhD
-Colony PCRed fliM, fliD, fliF
-Inoculated fliM
-PCRed flhD, flhB, flgJ, fliA to test directionality

8/27/12 week
-PCRed fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns, qseBC promoter, metR, araC, ompR
-Digested fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns, qseBC promoter, metR, araC, ompR
-all successful except ompR, metR, araC
-Digested pSB1C3
-Ligated fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns, qseBC promoter to pSB1C3

9/3/12 week
-Digested fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns
-Transformed BBa_K398108, BBa_J45199, BBa_K398014, BBa_J23039, BBa_K398406
-Digested pSB1C3
-Ligated fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns with pSB1C3

9/10/12 week
Transformed fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns
-Inoculated transformations
-Colony PCRed fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns
-Transformed BBa_K398108, BBa_J45199, BBa_K398014, BBa_J23039, BBa_K398406
-Colony PCRed BBa_K398108, BBa_J45199, BBa_K398014, BBa_J23039, BBa_K398406
-Transformed BBa_K398108, BBa_K398406
-Inoculated BBa_K398108, BBa_K398406, fliH
-Miniprepped BBa_K398108, BBa_K398406, fliH

-acceptable concentrations

-Colony PCRed BBa_K398108, BBa_K398406, fliH
-Miniprepped flhD

9/17/12 week
-Transformed arabinose
Inoculated cheY, T7
-Digested cheZ, cheZ RBS, cheZ, T7 Stp, cheY, 4-12A Stp, P.cola, pACYC
-Ligated cheZ, cheZ RBS, cheZ, T7 Stp, cheY, 4-12A Stp, P.cola, pACYC
-Miniprepped T7, cheY
-acceptable concentrations
-Transformed T7/p.cola, pACYC
-Colony PCRed cheZ RBS, cheZ T7, p.cola cheY
-Inoculated cheZ RBS, cheZ T7, p.cola cheY
-Miniprepped cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ
-Transformed cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ
-Inoculated cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ
-Imaged cheZ, cheZ T7, cheZ RBS T7, cheZ T7
-Glycerol stocks of cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ
-Imaging of pcola cheZ A, cheZ RBS T7, cheZ RBS T7, cheZ T7, pcdf cheY A
-Inoculated cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ

9/24/12 week
-Imaged BBa_K398108, cheZ RBS T7, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ
-Imaged cheZ RBS T7, cheZ T7, pcdf cheY, control
-Digested cheY p.cola, pcdf cheY, pcdf cheZ, pcola cheZ, cheZ T7 RBS
-Transformed T7 RBS cheZ
-Inoculated salt biobricks from frozen stocks
-Colony PCRed T7 RBS cheZ
-Miniprepped cheZ RBS T7

10/1/12 week

-Miniprepped cheA, cheB, cheY, cheZ, motA. motB, T7 RBS cheZ, T7 RBS cheY, flgJ, fliA, fliH, fliC-GFP, flhB, flhD

-Western blot for T7 RBS cheY, T7 RBS cheZ

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+		.reblog-post .wpcom-enhanced-excerpt address,
+		.reblog-post .wpcom-enhanced-excerpt li,
+		.reblog-post .wpcom-enhanced-excerpt h1,
+		.reblog-post .wpcom-enhanced-excerpt h2,
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+		.reblog-post .wpcom-enhanced-excerpt h4,
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+		.reblog-post .wpcom-enhanced-excerpt p { font-size: 100% !important; }
+		.reblog-post .wpcom-enhanced-excerpt blockquote,
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-{| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"
+		/* ]]> */
-!align="center"|[[Team:USC|Home]]
+		</style>
-!align="center"|[[Team:USC/Team|Team]]
-!align="center"|[https://igem.org/Team.cgi?year=2012&team_name=USC Official Team Profile]
+	<style type="text/css">
-!align="center"|[[Team:USC/Project|Project]]
+	/* <![CDATA[ */
-!align="center"|[[Team:USC/Parts|Parts Submitted to the Registry]]
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+		 * Link color
-!align="center"|[[Team:USC/Notebook|Notebook]]
+		 */
-!align="center"|[[Team:USC/Safety|Safety]]
+					a,
-!align="center"|[[Team:USC/Attributions|Attributions]]
+			.post-wrap a,
-|}
+			.by-author a,
+			#posts .post-content .post-foot a {
+				color: #e1c647;
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+				background-color: #bf1d2c;
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+			#access ul ul .current-menu-item > a,
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+				color: rgba( 255, 255, 255, 1 );
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+				background: rgba( 0, 0, 0, 0.3 );
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+			#access ul ul .current-menu-ancestor > a:hover,
+			#access ul ul .current_page_ancestor > a:hover {
+				background: rgba( 0, 0, 0, 0.3 );
+			}
+			/* ]]> */
+	</style>
+<meta name="application-name" content="E. musici" /><meta name="msapplication-window" content="width=device-width;height=device-height" /><meta name="msapplication-tooltip" content="University of Southern California iGEM 2012" /><meta name="msapplication-task" content="name=Subscribe;action-uri=http://uscigem2012.wordpress.com/feed/;icon-uri=http://s2.wp.com/i/favicon.ico" /><meta name="msapplication-task" content="name=Sign up for a free blog;action-uri=http://wordpress.com/signup/;icon-uri=http://s2.wp.com/i/favicon.ico" /><meta name="msapplication-task" content="name=WordPress.com Support;action-uri=http://support.wordpress.com/;icon-uri=http://s2.wp.com/i/favicon.ico" /><meta name="msapplication-task" content="name=WordPress.com Forums;action-uri=http://forums.wordpress.com/;icon-uri=http://s2.wp.com/i/favicon.ico" /><meta name="title" content="Lab&nbsp;Notebook | E. musici on WordPress.com" />
+<meta name="description" content="5/21/12 week -Transformed casBCED12 with Amp resistance, casBCED12 with Cm resistance, pSBIC3_RFP, pSBIK3_RFP, and pSBIA3_RFP             -successful results: pSBIC3_RFP and pSBIK3_RFP. -inoculated pSBIC3_RFP and pSBIK3_RFP -miniprepped pSBIK3_RFP, and pSBIT3_RFP             -all very low&hellip; Read More &rarr;" />
+		<style type="text/css">
+			#header-image {
+				background: url( 'http://uscigem2012.files.wordpress.com/2012/10/igem-header.gif' ) no-repeat;
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+				width: 960px;
+				height: 270px;
+			}
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+				display: block;
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+				height: 100%;
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+							#logo {
+					display: none;
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+body.custom-background { background-color: #ffffff; }
+</style>
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+	<div id="masthead">
+				<div class="container">
+			<div id="access">
+				<div class="menu"><ul><li class="page_item page-item-2"><a href="https://2012.igem.org/Team:USC">E.&nbsp;Musici</a></li><li class="page_item page-item-5"><a href="https://2012.igem.org/Team:USC/Team">Team</a></li><li class="page_item page-item-10"><a href="
+https://2012.igem.org/Team:USC/Project">Project</a></li><li class="page_item page-item-12"><a href="https://2012.igem.org/Team:USC/Parts">Our&nbsp;BioBricks</a></li><li class="page_item page-item-14 current_page_item"><a href="https://2012.igem.org/Team:USC/Notebook">Lab&nbsp;Notebook</a></li><li class="page_item page-item-16"><a href="https://2012.igem.org/Team:USC/Modeling">Human&nbsp;Practices</a></li><li class="page_item page-item-45"><a href="
+https://2012.igem.org/Team:USC/Safety">Safety</a></li></ul></div>
+			</div><!-- #access -->
+		</div><!-- .container -->
+	</div><!-- #masthead -->
-You should make use of the calendar feature on the wiki and start a lab notebook.  This may be looked at by the judges to see how your work progressed throughout the summer.  It is a very useful organizational tool as well.
+	<div id="header">
+		<div class="container">
+								<div id="header-image">
+						<a href="http://uscigem2012.wordpress.com/" title="E. musici" rel="home">E. musici</a>
+					</div><!-- #header-image -->
+						<div id="logo">
+				<h1>
+					<a href="http://uscigem2012.wordpress.com/">E. musici</a>
+				</h1>
+				<div class="desc">
+					University of Southern California iGEM 2012				</div><!-- .desc -->
+			</div><!-- #logo -->
+		</div><!-- .container -->
+	</div><!-- #header -->
+				<div id="post-14" class="post-14 page type-page status-publish hentry">
+					<div class="post-content">
+						<h2 class="postitle">
+							Lab&nbsp;Notebook						</h2><!-- .postitle -->
+						<p><span style="color:#000000;"><strong>5/21/12 week</strong></span><br />
+<span style="color:#000000;"> -Transformed casBCED12 with Amp resistance, casBCED12 with Cm resistance, pSBIC3_RFP, pSBIK3_RFP, and pSBIA3_RFP</span><br />
+<span style="color:#000000;">             -successful results: pSBIC3_RFP and pSBIK3_RFP.</span><br />
+<span style="color:#000000;"> -inoculated pSBIC3_RFP and pSBIK3_RFP</span><br />
+<span style="color:#000000;"> -miniprepped pSBIK3_RFP, and pSBIT3_RFP</span><br />
+<span style="color:#000000;">             -all very low concentration</span><br />
+<span style="color:#000000;"> -Miniprepped pSBIC3_RFP, pSBIA3_RFP</span></p>
+<p><span style="color:#000000;">-concentration (got 98 and 120 ug/ml, respectively)<a href="http://uscigem2012.files.wordpress.com/2012/07/dsc_0028.jpg"><img class="alignright size-medium wp-image-190" title="DSC_0028" src="http://uscigem2012.files.wordpress.com/2012/07/dsc_0028.jpg?w=442&h=292" alt="" width="442" height="292" /></a></span></p>
+<p><span style="color:#000000;">-Digested pSBIC3 from pSBIC3_RFP, pSBIK3 from pSBIK3_RFP</span><br />
+<span style="color:#000000;"> -Transformed pSBIT3_RFP, pSBIA3_RFP, casBCDE12 on Amp</span><br />
+<span style="color:#000000;"> -Digested casABCDE123, tetO-GFP</span><br />
+<span style="color:#000000;"> -Ligated casA,3 with tetO-GFP, with pSBIA3 or pSBIC3</span><br />
+<span style="color:#000000;"> -Ligated casBCDE12 with pSBIA3 or PSBIC3</span></p>
+<p><span style="color:#000000;"> <strong>5/28/12 week</strong></span><br />
+<span style="color:#000000;"> -Inoculated (2x) 5mL psB1A3_RFP and psB1T3_RFP.</span><br />
+<span style="color:#000000;"> -Transformed psB1C3_RFP, tetO_GFP, and psB3T5_RFP.</span><br />
+<span style="color:#000000;"> -Miniprepped psB1C3_RFP and psB1K3_RFP</span><br />
+<span style="color:#000000;"> -Transformed psB1C3-RFP, psB3T5-RFP, tetO-GFP</span><br />
+<span style="color:#000000;"> -PCRed fliE, fliM, flhD, flgA, fliD, flhB, fliA, fliE,fliF, fliG, fliH, flgJ, flgK, fliC, cheA, cheB, cheC, cheY, cheZ, luxS, lsrR, lsrK</span><br />
+<span style="color:#000000;"> -Gel extracted and digested the genes with EP or EX</span><br />
+<span style="color:#000000;"> -Miniprepped pSB3T5_RFP (3) and tetO-GFP</span><br />
+<span style="color:#000000;"> -Digested tetO-GFP 2</span></p>
+<p><span style="color:#000000;"> <strong>6/4/12 week</strong></span><br />
+<span style="color:#000000;"> -PCRed genes using BL21 and HT115</span><br />
+<span style="color:#000000;"> -Gel extracted and digested </span><br />
+<span style="color:#000000;"> -Ligated the genes to corresponding psb1C3_RFP vectors</span><br />
+<span style="color:#000000;"> -Transformed 1-9F, 1-14H, 1-18H, 3-2H, 3-2J, 3-8D, 4-12A</span><br />
+<span style="color:#000000;">             -successful: 4-12, 3-8D, 3-2H, 1-9F, 3-2J. (very low efficiency)</span><br />
+<span style="color:#000000;"> -Transformed cheB, cheC, cheY, cheZ, luxS, lsrK, lsrR, flaU, flbA, flhB, flhD, flgJ, flgK, fliA, fliC, fliD, fliF, fliG, fliH, fliM</span><br />
+<span style="color:#000000;"> -Inoculated 4-12A, 3-8D, 3-2H, 1-9F, 3-2J</span><br />
+<span style="color:#000000;"> -Inoculated HT115 (w/ 5uL tet) and HB101 (w/o antibiotics)</span><br />
+<span style="color:#000000;"> -PCRed fliE, fliM, flhD, flgA, fliD, flhB, fliA, fliE,fliF, fliG, fliH, flgJ, flgK, fliC, cheA, cheB, cheC, cheY, cheZ, luxS, lsrR, lsrK</span></p>
+<p><span style="color:#000000;"> <strong>6/11/12 week</strong></span><br />
+<span style="color:#000000;"> -Digested fliM, flhD, flgA, fliD, flhB, fliA, fliE,fliF, fliG, fliH, flgJ, flgK, fliC, cheA, cheB, cheC, cheY, cheZ, luxS, lsrR, lsrK</span><br />
+<span style="color:#000000;"> -Gel extracted fliM, flhD, flgA, fliD, flhB, fliA, fliE,fliF, fliG, fliH, flgJ, flgK, fliC, cheA, cheB, cheC, cheY, cheZ, luxS, lsrR, lsrK</span><br />
+<span style="color:#000000;"> -PCRed fliC with DH5alpha, BL21, HT115</span><br />
+<span style="color:#000000;"> -Ligated cm1, cm2, rfp1, rfp2</span><br />
+<span style="color:#000000;"> -Gel purified cm1, cm2, rfp1, rfp2</span><br />
+<span style="color:#000000;"> -Transformed fliA, cheZ, cheY, fliH, fliG, flgJ, flihB, flgK, fliD,fliM, 1-9F, 1-14H, 1-18H, 3-2H, 3-2J, 3-8D, 4-12A</span><br />
+<span style="color:#000000;">             -successful: 1-9F, 1-14H, 1-18H, 3-2H, 3-2J, 3-8D, 4-12A</span><br />
+<span style="color:#000000;"> -Digested cm vector with EP</span><br />
+<span style="color:#000000;"> -Gel extracted cm vectors</span><br />
+<span style="color:#000000;"> -Inoculated 3-8D, 1-9F, 4-12A</span><br />
+<span style="color:#000000;"> -PCRed chA, cheB, cheC, cheY, cheZ</span><br />
+<span style="color:#000000;"> -Transformed ligation products: fliG, flgJ, cheY, cheZ, fliD, flhB, fliM, fliH, flgK</span><br />
+<span style="color:#000000;"> -Ligated fliM, fliD, flhB, flgK, fliA, cheY, cheZ, fliG, flgJ, rfp1, rfp2 to Cm vector, fliH</span><br />
+<span style="color:#000000;"> -Miniprepped the innoculations (3-8D, 1-9F, 4-12A)</span><br />
+<span style="color:#000000;">             -low concentrations</span><br />
+<span style="color:#000000;"> -Transformed fliH, fliD, flhB, flgK, fliA, cheY, cheZ, fliG, flgJ, rfp1, rfp2, cm vector</span><br />
+<span style="color:#000000;"> -Miniprepped 1-9F(1), 1-9F(2), 3-8D(1), 3-8D(2), 4-12A(1), 4-12A(2)</span><br />
+<span style="color:#000000;">             -low concentrations</span></p>
+<p><span style="color:#000000;"><a href="http://uscigem2012.files.wordpress.com/2012/07/mg_4058.jpg"><img class="alignright size-medium wp-image-165" title="_MG_4058" src="http://uscigem2012.files.wordpress.com/2012/07/mg_4058.jpg?w=412&h=274" alt="" width="412" height="274" /></a><strong>6/18/12 week</strong></span><br />
+<span style="color:#000000;"> -PCRed lsrR(BL21), lsrR(HT115), lrsK(BL21), lsrK(HT115)</span><br />
+<span style="color:#000000;"> -Inoculated 1-9F, 3-8D, 4-12A</span><br />
+<span style="color:#000000;"> -Miniprepped 1-9F, 3-8D, 4-12A</span><br />
+<span style="color:#000000;">             -low concentrations</span><br />
+<span style="color:#000000;"> -Transformed J23100, RBS B0034, luxS, GFP E0040</span><br />
+<span style="color:#000000;">             -successful</span><br />
+<span style="color:#000000;"> -Inoculated J23100, RBS B0034, luxS, GFP E0040</span><br />
+<span style="color:#000000;"> -PCRed cheA, cheB, cheY, cheZ using HTB genomic DNA</span><br />
+<span style="color:#000000;"> -PCRed fliC(1), motA(2), motB(3) using DH5alpha</span><br />
+<span style="color:#000000;"> -Miniprepped 1-9F, 3-8D, 4-12A</span><br />
+<span style="color:#000000;">             -acceptable concentrations</span><br />
+<span style="color:#000000;"> -Digested cheA, cheB, cheY, cheZ and gel purified</span><br />
+<span style="color:#000000;"> -Ligated cheA, cheB, cheY, cheZ</span><br />
+<span style="color:#000000;"> -Transformed cheA, cheB, cheY, cheZ</span><br />
+<span style="color:#000000;">             -successful</span><br />
+<span style="color:#000000;"> -Digested psB1C3 with X and X/S</span><br />
+<span style="color:#000000;"> -Digested fliC, motA, motB</span><br />
+<span style="color:#000000;"> -Miniprepped J23100, GFP E0040, luxS, RBS Boo34</span><br />
+<span style="color:#000000;">             -low concentrations</span></p>
+<p><span style="color:#000000;"> <strong>6/25/12 week</strong></span><br />
+<span style="color:#000000;"> -Transformed motA, motB, and control</span><br />
+<span style="color:#000000;"> -Inoculated T7 promoter and cheY</span><br />
+<span style="color:#000000;"> -Colony PCRed 20 colonies from motA, motB</span><br />
+<span style="color:#000000;"> -Inoculated motA, motB, cheA, cheB, cheY, cheZ</span><br />
+<span style="color:#000000;"> -Miniprepped cheZ, cheY, cheA, cheB, motA, motB</span><br />
+<span style="color:#000000;">             -acceptable concentrations</span><br />
+<span style="color:#000000;"> -Glycerol stock: motA 6-14, cheZ, cheY, cheA, cheB, motA, motB</span><br />
+<span style="color:#000000;"> -Transformed cheZ, pLaeI+RBS+cheZ, RBS+cheZ+dbl terminator, cheZ w/ H13 tag+dbl terminator, cheZ w/ H13 tag, strong promoter w/RBS</span><br />
+<span style="color:#000000;"> -Inoculated cheZ, pLaeI+RBS+cheZ, RBS+cheZ+dbl terminator, cheZ w/ H13 tag+dbl terminator, cheZ w/ H13 tag, strong promoter w/RBS</span><br />
+<span style="color:#000000;"> -PCR miniprep of cheA(7,16), cheB(14), cheZ(14), motA(15)</span></p>
+<p><span style="color:#000000;"> <strong>7/2/12 week</strong></span><br />
+<span style="color:#000000;"> -Miniprepped cheZ w/ histag, cheZ, cheZ w/ RBS + dbl termination, pLacI w/ RBS + cheZ, cheZ w/ Histag + dbl termination, strong promoter w/ RBS (cm)</span><br />
+<span style="color:#000000;">             -acceptable concentrations</span><br />
+<span style="color:#000000;"> -Inoculated cheZ w/ histag (amp), cheZ (amp), cheZ w/ RBS, double termination (amp), pLacI w/ RBS, cheZ (amp), cheZ w/ histag, double termination (amp), strong promoter w/ RBS (cm)</span><br />
+<span style="color:#000000;"> -Miniprepped: psB1C3RFP (5 times), cheZ, strong promoter+RBS+cm, RBS+cheZ+dbl term site, pLacI+RBS+cheZ, cheZ+Hist+dbl, cheZ+Histag</span><br />
+<span style="color:#000000;">             -acceptable concentrations</span></p>
+<p><span style="color:#000000;"> <strong>7/9/12 week</strong></span><br />
+<span style="color:#000000;"> -Digested and gel extracted fliC and GFP</span><br />
+<span style="color:#000000;"> -Ligated fliC</span><br />
+<span style="color:#000000;"> -Transformed fliC-GFP</span><br />
+<span style="color:#000000;"> -Received and plated strains: BW30044, BW30045, K-12 wildtype strain, strain B wildtype, JW1871-1, JW1879-2, JW2662-1, MG1655 (motile), W3110           </span></p>
+<p><span style="color:#000000;"> <strong>7/16/12 week</strong></span><br />
+<span style="color:#000000;"> -Made competent cells MG1655     </span><br />
+<span style="color:#000000;"> -digested GFP vector for control using XbaI<a href="http://uscigem2012.files.wordpress.com/2012/07/dsc_0026.jpg"><img class="alignright size-medium wp-image-187" title="DSC_0026" src="http://uscigem2012.files.wordpress.com/2012/07/dsc_0026.jpg?w=512&h=338" alt="" width="512" height="338" /></a></span><br />
+<span style="color:#000000;"> -Colony PCRed the fliC GFP </span><br />
+<span style="color:#000000;">             -primer dimers. unsuccessful-</span></p>
+<p><span style="color:#000000;"> <strong>7/23/12 week</strong></span><br />
+<span style="color:#000000;"> -PCRed fliC w/ RBS</span><br />
+<span style="color:#000000;"> -Transformed GFP-DH5a, GFP-MG1655     </span><br />
+<span style="color:#000000;"> -Inoculated w/ 50uL transformation product</span><br />
+<span style="color:#000000;"> -Plated GFP-DH5a, GFP-MG1655</span><br />
+<span style="color:#000000;"> -PCRed fliC w/ RBS</span><br />
+<span style="color:#000000;"> -Miniprepped GFP-DH5a (x2), GFP-MG1655 (x2)</span></p>
+<p><span style="color:#000000;">-acceptable concentrations</span></p>
+<p><span style="color:#000000;">-Digested fliC and GFP w/ XbaI and gel purified</span><br />
+<span style="color:#000000;"> -Ligated fliC-GFP and GFP w/ H2O (control)</span><br />
+<span style="color:#000000;"> -Transformed 1hour ligation product</span><br />
+<span style="color:#000000;">             -unsuccessul</span><br />
+<span style="color:#000000;"> -Made control and cheY mutagenesis solutions</span><br />
+<span style="color:#000000;"> -Transformed fliC-GFP and GFP w/ H2O control into DH5a cells</span><br />
+<span style="color:#000000;">             -successful: GFP + H2O control and fliC-GFP</span><br />
+<span style="color:#000000;"> -Quickchange mutagenesis</span><br />
+<span style="color:#000000;"> -Colony PCR fliC-GFP</span></p>
+<p><span style="color:#000000;"> <strong>7/30/12 week</strong></span><br />
+<span style="color:#000000;"> -Miniprepped fliC-GFP</span><br />
+<span style="color:#000000;">             -acceptable concentrations</span><br />
+<span style="color:#000000;"> -Digested miniprepped products</span><br />
+<span style="color:#000000;"> -Quickchange PCRed fliC-GFP</span><br />
+<span style="color:#000000;"> -Digested quickchange PCR products with DPN1</span><br />
+<span style="color:#000000;"> -Digested fliC minipreps with Spe1 </span><br />
+<span style="color:#000000;">             -successful</span><br />
+<span style="color:#000000;"> -Transformed 43, 44, quickchange digests</span></p>
+<p><span style="color:#000000;"> <strong>8/6/12 week</strong></span><br />
+<span style="color:#000000;"> -Digested cheY</span><br />
+<span style="color:#000000;"> -Quickchange PCRed fliC and cheY with control</span><br />
+<span style="color:#000000;"> -Digested fliC-GFP, cheY, control</span><br />
+<span style="color:#000000;"> -Plated fliC, cheY, and pUC18</span><br />
+<span style="color:#000000;"> -Inoculated cheY mut colonies</span><br />
+<span style="color:#000000;"> -Miniprepped cheY mut</span><br />
+<span style="color:#000000;"> -Glycerol stocks of cheY mut</span><br />
+<span style="color:#000000;"> -PCRed cheY mut</span><br />
+<span style="color:#000000;"> -Digested cheY mut</span></p>
+<p><span style="color:#000000;"> <strong>8/13/12 week</strong></span><br />
+<span style="color:#000000;"> -PCRed fliA, fliD, fliE, fliF, fliG, fliH, fliM, flhB, flhC, flhD, flgA, flgB, flgJ, flgK, fliC-GFP, lsrR, lsrK</span><br />
+<span style="color:#000000;"> -Digested and gel extracted the flagella genes</span><br />
+<span style="color:#000000;"> -Transformed fliA, fliD,fliF, fliM, flhB, flgB, flgJ, flhD, lsrR, lsrK</span><br />
+<span style="color:#000000;"> -Digested fliC-GFP with DpnI, BBa_K318512, BBa_K338001, BBa_K216005</span><br />
+<span style="color:#000000;"> -Transformed pSB1C3 and fliC-GFP mut</span><br />
+<span style="color:#000000;"> -Inoculated pSB1C3-RFP, BBa_K318512, BBa_K338001, BBa_K216005</span><br />
+<span style="color:#000000;"> -Miniprepped pSB1C3 and BBa_K318512, BBa_K338001, BBa_K216005</span><br />
+<span style="color:#000000;">             -acceptable concentrations</span><br />
+<span style="color:#000000;"> -Digested pSB1C3 fliF, fliD, flhB, flhD, flgJ, fliM, fliA</span><br />
+<span style="color:#000000;"> -Ligated fliF, fliD, flhB, flhD, flgJ, fliM, fliA with pSB1C3</span></p>
+<p><span style="color:#000000;"> <strong>8/20/12 week</strong></span><br />
+<span style="color:#000000;"> -Colony PCRed fliA, fliM, fliD, fliF, flgJ, flhB, flhD</span><br />
+<span style="color:#000000;"> -Imaged MG1655 in various conditions: pH 5, 6, 8, 9; NaCl 50mM, 100mM, 200mM, 400mM, 500mM; temperature 15C, 20C, 42C; glucose 10mM, 20mM, 50mM</span><br />
+<span style="color:#000000;"> -Miniprepped fliC-GFP mut, cheY mut</span><br />
+<span style="color:#000000;">             -acceptable concentrations</span><br />
+<span style="color:#000000;"> -Glycerol stocks of fliC-GFP, cheY </span><br />
+<span style="color:#000000;"> -Colony PCRed flgJ, flhD, fliA, flhB, cheY</span><br />
+<span style="color:#000000;"> -Inoculated successful colonies</span><br />
+<span style="color:#000000;"> -Colony PCRed fliM, fliF, fliD</span><br />
+<span style="color:#000000;"> -Glycerol stocks of fliA, flhB, flgJ, flhD</span><br />
+<span style="color:#000000;"> -Imaging of MG1655</span><br />
+<span style="color:#000000;"> -Miniprepped fliA, flhB, flgJ, flhD</span><br />
+<span style="color:#000000;"> -Colony PCRed fliM, fliD, fliF</span><br />
+<span style="color:#000000;"> -Inoculated fliM </span><br />
+<span style="color:#000000;"> -PCRed flhD, flhB, flgJ, fliA to test directionality</span></p>
+<p><span style="color:#000000;"> <strong>8/27/12 week</strong></span><br />
+<span style="color:#000000;"> -PCRed fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns, qseBC promoter, metR, araC, ompR</span><br />
+<span style="color:#000000;"> -Digested fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns, qseBC promoter, metR, araC, ompR</span><br />
+<span style="color:#000000;">             -all successful except ompR, metR, araC</span><br />
+<span style="color:#000000;"> -Digested pSB1C3 </span><br />
+<span style="color:#000000;"> -Ligated fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns, qseBC promoter to pSB1C3</span></p>
+<p><span style="color:#000000;"> <strong>9/3/12 week</strong></span><br />
+<span style="color:#000000;"> -Digested  fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns </span><br />
+<span style="color:#000000;"> -Transformed BBa_K398108, BBa_J45199, BBa_K398014, BBa_J23039, BBa_K398406</span><br />
+<span style="color:#000000;"> -Digested pSB1C3</span><br />
+<span style="color:#000000;"> -Ligated  fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns with pSB1C3</span></p>
+<p><span style="color:#000000;"> <strong>9/10/12 week</strong></span><br />
+<span style="color:#000000;"> Transformed  fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns</span><br />
+<span style="color:#000000;"> -Inoculated transformations </span><br />
+<span style="color:#000000;"> -Colony PCRed  fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns</span><br />
+<span style="color:#000000;"> -Transformed BBa_K398108, BBa_J45199, BBa_K398014, BBa_J23039, BBa_K398406 </span><br />
+<span style="color:#000000;"> -Colony PCRed BBa_K398108, BBa_J45199, BBa_K398014, BBa_J23039, BBa_K398406</span><br />
+<span style="color:#000000;"> -Transformed BBa_K398108, BBa_K398406</span><br />
+<span style="color:#000000;"> -Inoculated BBa_K398108, BBa_K398406, fliH</span><br />
+<span style="color:#000000;"> -Miniprepped BBa_K398108, BBa_K398406, fliH</span></p>
+<p><span style="color:#000000;">-acceptable concentrations<a href="http://uscigem2012.files.wordpress.com/2012/07/mg_4057.jpg"><img class="alignright size-medium wp-image-162" title="_MG_4057" src="http://uscigem2012.files.wordpress.com/2012/07/mg_4057.jpg?w=496&h=331" alt="" width="496" height="331" /></a></span></p>
+<p><span style="color:#000000;">-Colony PCRed BBa_K398108, BBa_K398406, fliH           </span><br />
+<span style="color:#000000;"> -Miniprepped flhD</span></p>
+<p><span style="color:#000000;"> <strong>9/17/12 week</strong></span><br />
+<span style="color:#000000;"> -Transformed arabinose</span><br />
+<span style="color:#000000;"> Inoculated cheY, T7</span><br />
+<span style="color:#000000;"> -Digested cheZ, cheZ RBS, cheZ, T7 Stp, cheY, 4-12A Stp, P.cola, pACYC</span><br />
+<span style="color:#000000;"> -Ligated cheZ, cheZ RBS, cheZ, T7 Stp, cheY, 4-12A Stp, P.cola, pACYC</span><br />
+<span style="color:#000000;"> -Miniprepped T7, cheY</span><br />
+<span style="color:#000000;">             -acceptable concentrations</span><br />
+<span style="color:#000000;"> -Transformed T7/p.cola, pACYC</span><br />
+<span style="color:#000000;"> -Colony PCRed cheZ RBS, cheZ T7, p.cola cheY</span><br />
+<span style="color:#000000;"> -Inoculated cheZ RBS, cheZ T7, p.cola cheY</span><br />
+<span style="color:#000000;"> -Miniprepped cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ</span><br />
+<span style="color:#000000;"> -Transformed cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ</span><br />
+<span style="color:#000000;"> -Inoculated cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ</span><br />
+<span style="color:#000000;"> -Imaged cheZ, cheZ T7, cheZ RBS T7, cheZ T7</span><br />
+<span style="color:#000000;"> -Glycerol stocks of cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ</span><br />
+<span style="color:#000000;"> -Imaging of pcola cheZ A, cheZ RBS T7, cheZ RBS T7, cheZ T7, pcdf cheY A</span><br />
+<span style="color:#000000;"> -Inoculated cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ</span></p>
+<p><span style="color:#000000;"> <strong>9/24/12 week</strong></span><br />
+<span style="color:#000000;"> -Imaged BBa_K398108, cheZ RBS T7, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ</span><br />
+<span style="color:#000000;"> -Imaged cheZ RBS T7, cheZ T7, pcdf cheY, control</span><br />
+<span style="color:#000000;"> -Digested cheY p.cola, pcdf cheY, pcdf cheZ, pcola cheZ, cheZ T7 RBS</span><br />
+<span style="color:#000000;"> -Transformed T7 RBS cheZ </span><br />
+<span style="color:#000000;"> -Inoculated salt biobricks from frozen stocks</span><br />
+<span style="color:#000000;"> -Colony PCRed T7 RBS cheZ </span><br />
+<span style="color:#000000;"> -Miniprepped cheZ RBS T7</span></p>
+<p><span style="color:#000000;"> <strong>10/1/12 week</strong></span></p>
+<p><span style="color:#000000;">-Miniprepped cheA, cheB, cheY, cheZ, motA. motB, T7 RBS cheZ, T7 RBS cheY, flgJ, fliA, fliH, fliC-GFP, flhB, flhD</span></p>
+<p>-Western blot for T7 RBS cheY, T7 RBS cheZ</p>
+<div id="jp-post-flair" class="sharedaddy sd-like-enabled sd-sharing-enabled"></div>																	</div><!-- .post-content -->
+				</div><!-- #post-14 -->
+</body>
+</html>