Team:USC/Notebook

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Latest revision as of 03:05, 4 October 2012

Lab Notebook « E. musici

Lab Notebook

5/21/12 week
-Megan transformed casBCED12 with Amp resistance, casBCED12 with Cm resistance, pSBIC3_RFP, pSBIK3_RFP, and pSBIA3_RFP
-successful results: pSBIC3_RFP and pSBIK3_RFP.
-Luke inoculated pSBIC3_RFP and pSBIK3_RFP
-Stephan miniprepped pSBIK3_RFP, and pSBIT3_RFP
-all very low concentration
-Ellen miniprepped pSBIC3_RFP, pSBIA3_RFP

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

-Rachel digested pSBIC3 from pSBIC3_RFP, pSBIK3 from pSBIK3_RFP
-Eric transformed pSBIT3_RFP, pSBIA3_RFP, casBCDE12 on Amp
-Stephan digested casABCDE123, tetO-GFP
-Stephan digated casA,3 with tetO-GFP, with pSBIA3 or pSBIC3
-Eric digated casBCDE12 with pSBIA3 or PSBIC3

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

6/4/12 week
-Stephan PCRed genes using BL21 and HT115
-Stephan gel extracted and digested
-Ellen digested the genes to corresponding psb1C3_RFP vectors
-Rebecca 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)
-Rachel transformed cheB, cheC, cheY, cheZ, luxS, lsrK, lsrR, flaU, flbA, flhB, flhD, flgJ, flgK, fliA, fliC, fliD, fliF, fliG, fliH, fliM
-Rachel inoculated 4-12A, 3-8D, 3-2H, 1-9F, 3-2J
-Luke inoculated HT115 (w/ 5uL tet) and HB101 (w/o antibiotics)
-Stephan 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
-Ellen digested fliM, flhD, flgA, fliD, flhB, fliA, fliE,fliF, fliG, fliH, flgJ, flgK, fliC, cheA, cheB, cheC, cheY, cheZ, luxS, lsrR, lsrK
-Luke gel extracted fliM, flhD, flgA, fliD, flhB, fliA, fliE,fliF, fliG, fliH, flgJ, flgK, fliC, cheA, cheB, cheC, cheY, cheZ, luxS, lsrR, lsrK
-Rebecca PCRed fliC with DH5alpha, BL21, HT115
-Rachel ligated cm1, cm2, rfp1, rfp2
-Eric gel purified cm1, cm2, rfp1, rfp2
-Megan 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
-Stephan digested cm vector with EP
-Ellen gel extracted cm vectors
-Ellen inoculated 3-8D, 1-9F, 4-12A
-Eric PCRed chA, cheB, cheC, cheY, cheZ
-Luke transformed ligation products: fliG, flgJ, cheY, cheZ, fliD, flhB, fliM, fliH, flgK
-Rachel ligated fliM, fliD, flhB, flgK, fliA, cheY, cheZ, fliG, flgJ, rfp1, rfp2 to Cm vector, fliH
-Megan miniprepped the innoculations (3-8D, 1-9F, 4-12A)
-low concentrations
-Rebecca transformed fliH, fliD, flhB, flgK, fliA, cheY, cheZ, fliG, flgJ, rfp1, rfp2, cm vector
-Megan 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
-Rachel PCRed lsrR(BL21), lsrR(HT115), lrsK(BL21), lsrK(HT115)
-Stephan inoculated 1-9F, 3-8D, 4-12A
-Ellen miniprepped 1-9F, 3-8D, 4-12A
-low concentrations
-Luke transformed J23100, RBS B0034, luxS, GFP E0040
-successful
-Stephan inoculated J23100, RBS B0034, luxS, GFP E0040
-Stephan PCRed cheA, cheB, cheY, cheZ using HTB genomic DNA
-Stephan PCRed fliC(1), motA(2), motB(3) using DH5alpha
-Luke miniprepped 1-9F, 3-8D, 4-12A
-acceptable concentrations
-Megan digested cheA, cheB, cheY, cheZ and gel purified
-Luke ligated cheA, cheB, cheY, cheZ
-Megan transformed cheA, cheB, cheY, cheZ
-successful
-Eric digested psB1C3 with X and X/S
-Rachel digested fliC, motA, motB
-Rachel miniprepped J23100, GFP E0040, luxS, RBS Boo34
-low concentrations

6/25/12 week
-Ellen transformed motA, motB, and control
-Rebecca inoculated T7 promoter and cheY
-Rebecca colony PCRed 20 colonies from motA, motB
-Stephan inoculated motA, motB, cheA, cheB, cheY, cheZ
-Luke miniprepped cheZ, cheY, cheA, cheB, motA, motB
-acceptable concentrations
-Rachel glycerol stock: motA 6-14, cheZ, cheY, cheA, cheB, motA, motB
-Luke transformed cheZ, pLaeI+RBS+cheZ, RBS+cheZ+dbl terminator, cheZ w/ H13 tag+dbl terminator, cheZ w/ H13 tag, strong promoter w/RBS
-Rebecca inoculated cheZ, pLaeI+RBS+cheZ, RBS+cheZ+dbl terminator, cheZ w/ H13 tag+dbl terminator, cheZ w/ H13 tag, strong promoter w/RBS
-Stephan PCR miniprep of cheA(7,16), cheB(14), cheZ(14), motA(15)

7/2/12 week
-Megan 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
-Megan 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)
-Ellen 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
-Ellen digested and gel extracted fliC and GFP
-Rachel ligated fliC
-Luke transformed fliC-GFP
-Rebecca 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
-Rachel made competent cells MG1655
-Ellen digested GFP vector for control using XbaI
-Luke colony PCRed the fliC GFP
-primer dimers. unsuccessful-

7/23/12 week
-Ellen PCRed fliC w/ RBS
-Stephan transformed GFP-DH5a, GFP-MG1655
-Megan inoculated w/ 50uL transformation product
-Stephan plated GFP-DH5a, GFP-MG1655
-Rachel PCRed fliC w/ RBS
-Ellen miniprepped GFP-DH5a (x2), GFP-MG1655 (x2)

-acceptable concentrations

-Digested fliC and GFP w/ XbaI and gel purified
-Eric ligated fliC-GFP and GFP w/ H2O (control)
-Eric transformed 1hour ligation product
-unsuccessul
-Stephan made control and cheY mutagenesis solutions
-Megan transformed fliC-GFP and GFP w/ H2O control into DH5a cells
-successful: GFP + H2O control and fliC-GFP
-Luke quickchange mutagenesis
-Rebecca colony PCR fliC-GFP

7/30/12 week
-Megan miniprepped fliC-GFP
-acceptable concentrations
-Rachel digested miniprepped products
-Eric quickchange PCRed fliC-GFP
-Stephan digested quickchange PCR products with DPN1
-Megan digested fliC minipreps with Spe1
-successful
-Megan transformed 43, 44, quickchange digests

8/6/12 week
-Rebecca digested cheY
-Rebecca quickchange PCRed fliC and cheY with control
-Rebecca digested fliC-GFP, cheY, control
-Stephan plated fliC, cheY, and pUC18
-Rachel inoculated cheY mut colonies
-Ellen miniprepped cheY mut
-Stephan glycerol stocks of cheY mut
-Luke PCRed cheY mut
-Luke 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
-Ellen digested and gel extracted the flagella genes
-Stephan transformed fliA, fliD,fliF, fliM, flhB, flgB, flgJ, flhD, lsrR, lsrK
-Megan digested fliC-GFP with DpnI, BBa_K318512, BBa_K338001, BBa_K216005
-Rebecca transformed pSB1C3 and fliC-GFP mut
-Stephan inoculated pSB1C3-RFP, BBa_K318512, BBa_K338001, BBa_K216005
-Ellen miniprepped pSB1C3 and BBa_K318512, BBa_K338001, BBa_K216005
-acceptable concentrations
-Luke digested pSB1C3 fliF, fliD, flhB, flhD, flgJ, fliM, fliA
-Eric ligated fliF, fliD, flhB, flhD, flgJ, fliM, fliA with pSB1C3

8/20/12 week
-Megan colony PCRed fliA, fliM, fliD, fliF, flgJ, flhB, flhD
-Rachel imaged MG1655 in various conditions: pH 5, 6, 8, 9; NaCl 50mM, 100mM, 200mM, 400mM, 500mM; temperature 15C, 20C, 42C; glucose 10mM, 20mM, 50mM
-Rachel miniprepped fliC-GFP mut, cheY mut
-acceptable concentrations
-Stephan glycerol stocks of fliC-GFP, cheY
-Megan colony PCRed flgJ, flhD, fliA, flhB, cheY
-Megan inoculated successful colonies
-Rachel colony PCRed fliM, fliF, fliD
-Rebecca glycerol stocks of fliA, flhB, flgJ, flhD
-Eric imaging of MG1655
-Luke miniprepped fliA, flhB, flgJ, flhD
-Luke colony PCRed fliM, fliD, fliF
-Rebecca inoculated fliM
-Stephan PCRed flhD, flhB, flgJ, fliA to test directionality

8/27/12 week
-Ellen PCRed fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns, qseBC promoter, metR, araC, ompR
-Eric 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
-Stephan digested pSB1C3
-Megan 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
-Rachel digested fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns
-Eric transformed BBa_K398108, BBa_J45199, BBa_K398014, BBa_J23039, BBa_K398406
-Stephan digested pSB1C3
-Luke ligated fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns with pSB1C3

9/10/12 week
-Megan transformed fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns
-Stephan inoculated transformations
-Rebecca colony PCRed fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns
-Ellen transformed BBa_K398108, BBa_J45199, BBa_K398014, BBa_J23039, BBa_K398406
-Megan colony PCRed BBa_K398108, BBa_J45199, BBa_K398014, BBa_J23039, BBa_K398406
-Rachel transformed BBa_K398108, BBa_K398406
-Luke inoculated BBa_K398108, BBa_K398406, fliH
-Eric miniprepped BBa_K398108, BBa_K398406, fliH

-acceptable concentrations

-Eric colony PCRed BBa_K398108, BBa_K398406, fliH
-Rachel miniprepped flhD

9/17/12 week
-Stephan transformed arabinose
-Stephan inoculated cheY, T7
-Rachel digested cheZ, cheZ RBS, cheZ, T7 Stp, cheY, 4-12A Stp, P.cola, pACYC
-Luke ligated cheZ, cheZ RBS, cheZ, T7 Stp, cheY, 4-12A Stp, P.cola, pACYC
-Megan miniprepped T7, cheY
-acceptable concentrations
-Rebecca transformed T7/p.cola, pACYC
-Rebecca colony PCRed cheZ RBS, cheZ T7, p.cola cheY
-Rebecca inoculated cheZ RBS, cheZ T7, p.cola cheY
-Stephan miniprepped cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ
-Megan transformed cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ
-Sean inoculated cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ
-Megan imaged cheZ, cheZ T7, cheZ RBS T7, cheZ T7
-Ellen glycerol stocks of cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ
-Eric imaging of pcola cheZ A, cheZ RBS T7, cheZ RBS T7, cheZ T7, pcdf cheY A
-Luke inoculated cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ

9/24/12 week
-Rebecca imaged BBa_K398108, cheZ RBS T7, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ
-Stephan imaged cheZ RBS T7, cheZ T7, pcdf cheY, control
-Luke digested cheY p.cola, pcdf cheY, pcdf cheZ, pcola cheZ, cheZ T7 RBS
-Megan transformed T7 RBS cheZ
-Rebecca inoculated salt biobricks from frozen stocks
-Rachel colony PCRed T7 RBS cheZ
-Ellen miniprepped cheZ RBS T7

10/1/12 week

-Eric 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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+<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;" />
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-!align="center"|[[Team:USC|Home]]
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-!align="center"|[[Team:USC/Project|Project]]
+</style>
-!align="center"|[[Team:USC/Parts|Parts Submitted to the Registry]]
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-!align="center"|[[Team:USC/Modeling|Modeling]]
+<body class="page page-id-14 page-template-default custom-background single-author one-column gecko highlander-enabled highlander-light">
-!align="center"|[[Team:USC/Notebook|Notebook]]
-!align="center"|[[Team:USC/Safety|Safety]]
-!align="center"|[[Team:USC/Attributions|Attributions]]
-|}
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+				<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/HumanPractices">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>
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+					University of Southern California iGEM 2012				</div><!-- .desc -->
+			</div><!-- #logo -->
+		</div><!-- .container -->
+	</div><!-- #header -->
-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="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;"> -Megan 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;"> -Luke inoculated pSBIC3_RFP and pSBIK3_RFP</span><br />
+<span style="color:#000000;"> -Stephan miniprepped pSBIK3_RFP, and pSBIT3_RFP</span><br />
+<span style="color:#000000;">             -all very low concentration</span><br />
+<span style="color:#000000;"> -Ellen 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;">-Rachel digested pSBIC3 from pSBIC3_RFP, pSBIK3 from pSBIK3_RFP</span><br />
+<span style="color:#000000;"> -Eric transformed pSBIT3_RFP, pSBIA3_RFP, casBCDE12 on Amp</span><br />
+<span style="color:#000000;"> -Stephan digested <em>casABCDE123</em>, tetO-GFP</span><br />
+<span style="color:#000000;"> -Stephan digated <em>casA</em>,3 with tetO-GFP, with pSBIA3 or pSBIC3</span><br />
+<span style="color:#000000;"> -Eric digated casBCDE12 with pSBIA3 or PSBIC3</span></p>
+<p><span style="color:#000000;"> <strong>5/28/12 week</strong></span><br />
+<span style="color:#000000;"> -Megan inoculated (2x) 5mL psB1A3_RFP and psB1T3_RFP.</span><br />
+<span style="color:#000000;"> -Rebecca transformed psB1C3_RFP, tetO_GFP, and psB3T5_RFP.</span><br />
+<span style="color:#000000;"> -Rebecca miniprepped psB1C3_RFP and psB1K3_RFP</span><br />
+<span style="color:#000000;"> -Ellen transformed psB1C3-RFP, psB3T5-RFP, tetO-GFP</span><br />
+<span style="color:#000000;"> -Rachel PCRed <em>fliE, fliM, flhD, flgA, fliD, flhB, fliA, fliE,fliF, fliG, fliH, flgJ, flgK, fliC, cheA, cheB, cheC, cheY, cheZ, luxS, lsrR, lsrK</em></span><br />
+<span style="color:#000000;"> -Luke gel extracted and digested the genes with EP or EX</span><br />
+<span style="color:#000000;"> -Megan miniprepped pSB3T5_RFP (3) and tetO-GFP</span><br />
+<span style="color:#000000;"> -Rachel digested tetO-GFP 2</span></p>
+<p><span style="color:#000000;"> <strong>6/4/12 week</strong></span><br />
+<span style="color:#000000;"> -Stephan PCRed genes using BL21 and HT115</span><br />
+<span style="color:#000000;"> -Stephan gel extracted and digested </span><br />
+<span style="color:#000000;"> -Ellen digested the genes to corresponding psb1C3_RFP vectors</span><br />
+<span style="color:#000000;"> -Rebecca 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;"> -Rachel transformed <em>cheB, cheC, cheY, cheZ, luxS, lsrK, lsrR, flaU, flbA, flhB, flhD, flgJ, flgK, fliA, fliC, fliD, fliF, fliG, fliH, fliM</em></span><br />
+<span style="color:#000000;"> -Rachel inoculated 4-12A, 3-8D, 3-2H, 1-9F, 3-2J</span><br />
+<span style="color:#000000;"> -Luke inoculated HT115 (w/ 5uL tet) and HB101 (w/o antibiotics)</span><br />
+<span style="color:#000000;"> -Stephan PCRed <em>fliE, fliM, flhD, flgA, fliD, flhB, fliA, fliE,fliF, fliG, fliH, flgJ, flgK, fliC, cheA, cheB, cheC, cheY, cheZ, luxS, lsrR, lsrK</em></span></p>
+<p><span style="color:#000000;"> <strong>6/11/12 week</strong></span><br />
+<span style="color:#000000;"> -Ellen digested <em>fliM, flhD, flgA, fliD, flhB, fliA, fliE,fliF, fliG, fliH, flgJ, flgK, fliC, cheA, cheB, cheC, cheY, cheZ, luxS, lsrR, lsrK</em></span><br />
+<span style="color:#000000;"> -Luke gel extracted <em>fliM, flhD, flgA, fliD, flhB, fliA, fliE,fliF, fliG, fliH, flgJ, flgK, fliC, cheA, cheB, cheC, cheY, cheZ, luxS, lsrR, lsrK</em></span><br />
+<span style="color:#000000;"> -Rebecca PCRed <em>fliC</em> with DH5alpha, BL21, HT115</span><br />
+<span style="color:#000000;"> -Rachel ligated cm1, cm2, rfp1, rfp2</span><br />
+<span style="color:#000000;"> -Eric gel purified cm1, cm2, rfp1, rfp2</span><br />
+<span style="color:#000000;"> -Megan transformed <em>fliA, cheZ, cheY, fliH, fliG, flgJ, flihB, flgK, fliD,fliM</em>, 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;"> -Stephan digested cm vector with EP</span><br />
+<span style="color:#000000;"> -Ellen gel extracted cm vectors</span><br />
+<span style="color:#000000;"> -Ellen inoculated 3-8D, 1-9F, 4-12A</span><br />
+<span style="color:#000000;"> -Eric PCRed <em>chA, cheB, cheC, cheY, cheZ</em></span><br />
+<span style="color:#000000;"> -Luke transformed ligation products: <em>fliG, flgJ, cheY, cheZ, fliD, flhB, fliM, fliH, flgK</em></span><br />
+<span style="color:#000000;"> -Rachel ligated <em>fliM, fliD, flhB, flgK, fliA, cheY, cheZ, fliG, flgJ, rfp1, rfp2 to Cm vector, fliH</em></span><br />
+<span style="color:#000000;"> -Megan miniprepped the innoculations (3-8D, 1-9F, 4-12A)</span><br />
+<span style="color:#000000;">             -low concentrations</span><br />
+<span style="color:#000000;"> -Rebecca transformed <em>fliH, fliD, flhB, flgK, fliA, cheY, cheZ, fliG, flgJ, rfp1, rfp2</em>, cm vector</span><br />
+<span style="color:#000000;"> -Megan 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;"> -Rachel PCRed <em>lsrR</em>(BL21), <em>lsrR</em>(HT115), <em>lrsK</em>(BL21), <em>lsrK</em>(HT115)</span><br />
+<span style="color:#000000;"> -Stephan inoculated 1-9F, 3-8D, 4-12A</span><br />
+<span style="color:#000000;"> -Ellen miniprepped 1-9F, 3-8D, 4-12A</span><br />
+<span style="color:#000000;">             -low concentrations</span><br />
+<span style="color:#000000;"> -Luke transformed J23100, RBS B0034, <em>luxS</em>, GFP E0040</span><br />
+<span style="color:#000000;">             -successful</span><br />
+<span style="color:#000000;"> -Stephan inoculated J23100, RBS B0034, <em>luxS</em>, GFP E0040</span><br />
+<span style="color:#000000;"> -Stephan PCRed <em>cheA, cheB, cheY, cheZ</em> using HTB genomic DNA</span><br />
+<span style="color:#000000;"> -Stephan PCRed <em>fliC</em>(1), <em>motA</em>(2), <em>motB</em>(3) using DH5alpha</span><br />
+<span style="color:#000000;"> -Luke miniprepped 1-9F, 3-8D, 4-12A</span><br />
+<span style="color:#000000;">             -acceptable concentrations</span><br />
+<span style="color:#000000;"> -Megan digested <em>cheA, cheB, cheY, cheZ</em> and gel purified</span><br />
+<span style="color:#000000;"> -Luke ligated <em>cheA, cheB, cheY, cheZ</em></span><br />
+<span style="color:#000000;"> -Megan transformed <em>cheA, cheB, cheY, cheZ</em></span><br />
+<span style="color:#000000;">             -successful</span><br />
+<span style="color:#000000;"> -Eric digested psB1C3 with X and X/S</span><br />
+<span style="color:#000000;"> -Rachel digested <em>fliC, motA, motB</em></span><br />
+<span style="color:#000000;"> -Rachel miniprepped J23100, GFP E0040, <em>luxS</em>, 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;"> -Ellen transformed <em>motA, motB</em>, and control</span><br />
+<span style="color:#000000;"> -Rebecca inoculated T7 promoter and <em>cheY</em></span><br />
+<span style="color:#000000;"> -Rebecca colony PCRed 20 colonies from <em>motA, motB</em></span><br />
+<span style="color:#000000;"> -Stephan inoculated <em>motA, motB, cheA, cheB, cheY, cheZ</em></span><br />
+<span style="color:#000000;"> -Luke miniprepped <em>cheZ, cheY, cheA, cheB, motA, motB</em></span><br />
+<span style="color:#000000;">             -acceptable concentrations</span><br />
+<span style="color:#000000;"> -Rachel glycerol stock: <em>motA</em> 6-14, <em>cheZ, cheY, cheA, cheB, motA, motB</em></span><br />
+<span style="color:#000000;"> -Luke transformed <em>cheZ</em>, pLaeI+RBS+<em>cheZ</em>, RBS+<em>cheZ</em>+dbl terminator, <em>cheZ</em> w/ H13 tag+dbl terminator, <em>cheZ</em> w/ H13 tag, strong promoter w/RBS</span><br />
+<span style="color:#000000;"> -Rebecca inoculated <em>cheZ</em>, pLaeI+RBS+<em>cheZ</em>, RBS+<em>cheZ</em>+dbl terminator, <em>cheZ</em> w/ H13 tag+dbl terminator, <em>cheZ</em> w/ H13 tag, strong promoter w/RBS</span><br />
+<span style="color:#000000;"> -Stephan PCR miniprep of <em>cheA</em>(7,16), <em>cheB</em>(14), <em>cheZ</em>(14), <em>motA</em>(15)</span></p>
+<p><span style="color:#000000;"> <strong>7/2/12 week</strong></span><br />
+<span style="color:#000000;"> -Megan miniprepped <em>cheZ</em> w/ histag, <em>cheZ</em>, <em>cheZ</em> w/ RBS + dbl termination, pLacI w/ RBS + <em>cheZ</em>, <em>cheZ</em> w/ Histag + dbl termination, strong promoter w/ RBS (cm)</span><br />
+<span style="color:#000000;">             -acceptable concentrations</span><br />
+<span style="color:#000000;"> -Megan inoculated <em>cheZ</em> w/ histag (amp), <em>cheZ</em> (amp), <em>cheZ</em> w/ RBS, double termination (amp), pLacI w/ RBS, <em>cheZ</em> (amp), <em>cheZ</em> w/ histag, double termination (amp), strong promoter w/ RBS (cm)</span><br />
+<span style="color:#000000;"> -Ellen miniprepped: psB1C3RFP (5 times), <em>cheZ</em>, strong promoter+RBS+cm, RBS+<em>cheZ</em>+dbl term site, pLacI+RBS+<em>cheZ</em>, <em>cheZ</em>+Hist+dbl, <em>cheZ</em>+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;"> -Ellen digested and gel extracted <em>fliC</em> and <em>GFP</em></span><br />
+<span style="color:#000000;"> -Rachel ligated <em>fliC</em></span><br />
+<span style="color:#000000;"> -Luke transformed <em>fliC-GFP</em></span><br />
+<span style="color:#000000;"> -Rebecca 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;"> -Rachel made competent cells MG1655     </span><br />
+<span style="color:#000000;"> -Ellen 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;"> -Luke colony PCRed the <em>fliC GFP</em> </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;"> -Ellen PCRed <em>fliC w/ RBS</em></span><br />
+<span style="color:#000000;"> -Stephan transformed <em>GFP-DH5a, GFP-MG1655</em>     </span><br />
+<span style="color:#000000;"> -Megan inoculated w/ 50uL transformation product</span><br />
+<span style="color:#000000;"> -Stephan plated <em>GFP-DH5a, GFP-MG1655</em></span><br />
+<span style="color:#000000;"> -Rachel PCRed <em>fliC</em> w/ RBS</span><br />
+<span style="color:#000000;"> -Ellen miniprepped <em>GFP-DH5a (x2), GFP-MG1655 </em>(x2)</span></p>
+<p><span style="color:#000000;">-acceptable concentrations</span></p>
+<p><span style="color:#000000;">-Digested <em>fliC</em> and GFP w/ XbaI and gel purified</span><br />
+<span style="color:#000000;"> -Eric ligated <em>fliC-GFP</em> and GFP w/ H2O (control)</span><br />
+<span style="color:#000000;"> -Eric transformed 1hour ligation product</span><br />
+<span style="color:#000000;">             -unsuccessul</span><br />
+<span style="color:#000000;"> -Stephan made control and <em>cheY</em> mutagenesis solutions</span><br />
+<span style="color:#000000;"> -Megan transformed <em>fliC-GFP</em> and GFP w/ H2O control into DH5a cells</span><br />
+<span style="color:#000000;">             -successful: GFP + H2O control and <em>fliC-GFP</em></span><br />
+<span style="color:#000000;"> -Luke quickchange mutagenesis</span><br />
+<span style="color:#000000;"> -Rebecca colony PCR <em>fliC-GFP</em></span></p>
+<p><span style="color:#000000;"> <strong>7/30/12 week</strong></span><br />
+<span style="color:#000000;"> -Megan miniprepped <em>fliC-GFP</em></span><br />
+<span style="color:#000000;">             -acceptable concentrations</span><br />
+<span style="color:#000000;"> -Rachel digested miniprepped products</span><br />
+<span style="color:#000000;"> -Eric quickchange PCRed <em>fliC-GFP</em></span><br />
+<span style="color:#000000;"> -Stephan digested quickchange PCR products with DPN1</span><br />
+<span style="color:#000000;"> -Megan digested <em>fliC</em> minipreps with Spe1 </span><br />
+<span style="color:#000000;">             -successful</span><br />
+<span style="color:#000000;"> -Megan transformed 43, 44, quickchange digests</span></p>
+<p><span style="color:#000000;"> <strong>8/6/12 week</strong></span><br />
+<span style="color:#000000;"> -Rebecca digested <em>cheY</em></span><br />
+<span style="color:#000000;"> -Rebecca quickchange PCRed <em>fliC and cheY</em> with control</span><br />
+<span style="color:#000000;"> -Rebecca digested <em>fliC-GFP, cheY</em>, control</span><br />
+<span style="color:#000000;"> -Stephan plated <em>fliC, cheY, and pUC18</em></span><br />
+<span style="color:#000000;"> -Rachel inoculated <em>cheY</em> mut colonies</span><br />
+<span style="color:#000000;"> -Ellen miniprepped <em>cheY mut</em></span><br />
+<span style="color:#000000;"> -Stephan glycerol stocks of <em>cheY mut</em></span><br />
+<span style="color:#000000;"> -Luke PCRed <em>cheY</em> mut</span><br />
+<span style="color:#000000;"> -Luke digested <em>cheY</em> mut</span></p>
+<p><span style="color:#000000;"> <strong>8/13/12 week</strong></span><br />
+<span style="color:#000000;"> -PCRed <em>fliA, fliD, fliE, fliF, fliG, fliH, fliM, flhB, flhC, flhD, flgA, flgB, flgJ, flgK, fliC-GFP, lsrR, lsrK</em></span><br />
+<span style="color:#000000;"> -Ellen digested and gel extracted the flagella genes</span><br />
+<span style="color:#000000;"> -Stephan transformed <em>fliA, fliD,fliF, fliM, flhB, flgB, flgJ, flhD, lsrR, lsrK</em></span><br />
+<span style="color:#000000;"> -Megan digested <em>fliC-GFP</em> with DpnI, BBa_K318512, BBa_K338001, BBa_K216005</span><br />
+<span style="color:#000000;"> -Rebecca transformed pSB1C3 and <em>fliC-GFP</em> mut</span><br />
+<span style="color:#000000;"> -Stephan inoculated pSB1C3-RFP, BBa_K318512, BBa_K338001, BBa_K216005</span><br />
+<span style="color:#000000;"> -Ellen miniprepped pSB1C3 and BBa_K318512, BBa_K338001, BBa_K216005</span><br />
+<span style="color:#000000;">             -acceptable concentrations</span><br />
+<span style="color:#000000;"> -Luke digested pSB1C3 <em>fliF, fliD, flhB, flhD, flgJ, fliM, fliA</em></span><br />
+<span style="color:#000000;"> -Eric ligated <em>fliF, fliD, flhB, flhD, flgJ, fliM, fliA</em> with pSB1C3</span></p>
+<p><span style="color:#000000;"> <strong>8/20/12 week</strong></span><br />
+<span style="color:#000000;"> -Megan colony PCRed <em>fliA, fliM, fliD, fliF, flgJ, flhB, flhD</em></span><br />
+<span style="color:#000000;"> -Rachel 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;"> -Rachel miniprepped <em>fliC-GFP mut, cheY mut</em></span><br />
+<span style="color:#000000;">             -acceptable concentrations</span><br />
+<span style="color:#000000;"> -Stephan glycerol stocks of <em>fliC-GFP, cheY</em> </span><br />
+<span style="color:#000000;"> -Megan colony PCRed <em>flgJ, flhD, fliA, flhB, cheY</em></span><br />
+<span style="color:#000000;"> -Megan inoculated successful colonies</span><br />
+<span style="color:#000000;"> -Rachel colony PCRed <em>fliM, fliF, fliD</em></span><br />
+<span style="color:#000000;"> -Rebecca glycerol stocks of <em>fliA, flhB, flgJ, flhD</em></span><br />
+<span style="color:#000000;"> -Eric imaging of MG1655</span><br />
+<span style="color:#000000;"> -Luke miniprepped <em>fliA, flhB, flgJ, flhD</em></span><br />
+<span style="color:#000000;"> -Luke colony PCRed <em>fliM, fliD, fliF</em></span><br />
+<span style="color:#000000;"> -Rebecca inoculated <em>fliM</em> </span><br />
+<span style="color:#000000;"> -Stephan PCRed <em>flhD, flhB, flgJ, fliA</em> to test directionality</span></p>
+<p><span style="color:#000000;"> <strong>8/27/12 week</strong></span><br />
+<span style="color:#000000;"> -Ellen PCRed <em>fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns, qseBC promoter, metR, araC, ompR</em></span><br />
+<span style="color:#000000;"> -Eric digested <em>fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns, qseBC promoter, metR, araC, ompR</em></span><br />
+<span style="color:#000000;">             -all successful except <em>ompR, metR, araC</em></span><br />
+<span style="color:#000000;"> -Stephan digested pSB1C3 </span><br />
+<span style="color:#000000;"> -Megan ligated <em>fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns, qseBC</em> promoter to pSB1C3</span></p>
+<p><span style="color:#000000;"> <strong>9/3/12 week</strong></span><br />
+<span style="color:#000000;"> -Rachel digested  <em>fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns </em></span><br />
+<span style="color:#000000;"> -Eric transformed BBa_K398108, BBa_J45199, BBa_K398014, BBa_J23039, BBa_K398406</span><br />
+<span style="color:#000000;"> -Stephan digested pSB1C3</span><br />
+<span style="color:#000000;"> -Luke ligated  <em>fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns</em> with pSB1C3</span></p>
+<p><span style="color:#000000;"> <strong>9/10/12 week</strong></span><br />
+<span style="color:#000000;"> -Megan transformed  <em>fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns</em></span><br />
+<span style="color:#000000;"> -Stephan inoculated transformations </span><br />
+<span style="color:#000000;"> -Rebecca colony PCRed  <em>fliA, fliD, fliE, fliG, fliH, fliM, flhC, flhD, qseB, qseC, qseBC, flgA, flgB, flgJ, flgK, hns</em></span><br />
+<span style="color:#000000;"> -Ellen transformed BBa_K398108, BBa_J45199, BBa_K398014, BBa_J23039, BBa_K398406 </span><br />
+<span style="color:#000000;"> -Megan colony PCRed BBa_K398108, BBa_J45199, BBa_K398014, BBa_J23039, BBa_K398406</span><br />
+<span style="color:#000000;"> -Rachel transformed BBa_K398108, BBa_K398406</span><br />
+<span style="color:#000000;"> -Luke inoculated BBa_K398108, BBa_K398406, <em>fliH</em></span><br />
+<span style="color:#000000;"> -Eric miniprepped BBa_K398108, BBa_K398406, <em>fliH</em></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;">-Eric colony PCRed BBa_K398108, BBa_K398406, fliH           </span><br />
+<span style="color:#000000;"> -Rachel miniprepped <em>flhD</em></span></p>
+<p><span style="color:#000000;"> <strong>9/17/12 week</strong></span><br />
+<span style="color:#000000;"> -Stephan transformed arabinose</span><br />
+<span style="color:#000000;"> -Stephan inoculated <em>cheY</em>, T7</span><br />
+<span style="color:#000000;"> -Rachel digested <em>cheZ, cheZ RBS, cheZ, T7 Stp, cheY</em>, 4-12A Stp, P.cola, pACYC</span><br />
+<span style="color:#000000;"> -Luke ligated <em>cheZ, cheZ RBS, cheZ, T7 Stp, cheY</em>, 4-12A Stp, P.cola, pACYC</span><br />
+<span style="color:#000000;"> -Megan miniprepped T7, <em>cheY</em></span><br />
+<span style="color:#000000;">             -acceptable concentrations</span><br />
+<span style="color:#000000;"> -Rebecca transformed T7/p.cola, pACYC</span><br />
+<span style="color:#000000;"> -Rebecca colony PCRed <em>cheZ RBS, cheZ T7, p.cola cheY</em></span><br />
+<span style="color:#000000;"> -Rebecca inoculated <em>cheZ RBS, cheZ T7, p.cola cheY</em></span><br />
+<span style="color:#000000;"> -Stephan miniprepped <em>cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ</em></span><br />
+<span style="color:#000000;"> -Megan transformed <em>cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ</em></span><br />
+<span style="color:#000000;"> -Sean inoculated <em>cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ</em></span><br />
+<span style="color:#000000;"> -Megan imaged <em>cheZ, cheZ T7, cheZ RBS T7, cheZ T7</em></span><br />
+<span style="color:#000000;"> -Ellen glycerol stocks of <em>cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ</em></span><br />
+<span style="color:#000000;"> -Eric imaging of <em>pcola cheZ A, cheZ RBS T7, cheZ RBS T7, cheZ T7, pcdf cheY A</em></span><br />
+<span style="color:#000000;"> -Luke inoculated <em>cheZ RBS, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ</em></span></p>
+<p><span style="color:#000000;"> <strong>9/24/12 week</strong></span><br />
+<span style="color:#000000;"> -Rebecca imaged BBa_K398108, <em>cheZ RBS T7, cheZ T7, p.cola cheY, p.cola cheZ, pcdf cheY, pcdf cheZ</em></span><br />
+<span style="color:#000000;"> -Stephan imaged <em>cheZ RBS T7, cheZ T7, pcdf cheY</em>, control</span><br />
+<span style="color:#000000;"> -Luke digested <em>cheY p.cola, pcdf cheY, pcdf cheZ, pcola cheZ, cheZ T7 RBS</em></span><br />
+<span style="color:#000000;"> -Megan transformed <em>T7 RBS cheZ</em> </span><br />
+<span style="color:#000000;"> -Rebecca inoculated salt biobricks from frozen stocks</span><br />
+<span style="color:#000000;"> -Rachel colony PCRed <em>T7 RBS cheZ</em> </span><br />
+<span style="color:#000000;"> -Ellen miniprepped <em>cheZ RBS T7</em></span></p>
+<p><span style="color:#000000;"> <strong>10/1/12 week</strong></span></p>
+<p><span style="color:#000000;">-Eric miniprepped <em>cheA, cheB, cheY, cheZ, motA. motB, T7 RBS cheZ, T7 RBS cheY, flgJ, fliA, fliH, fliC-GFP, flhB, flhD</em></span></p>
+<p>-Western blot for <em>T7 RBS cheY, T7 RBS cheZ</em></p>
+<div id="jp-post-flair" class="sharedaddy sd-like-enabled sd-sharing-enabled"></div>																	</div><!-- .post-content -->
+				</div><!-- #post-14 -->
+</body>
+</html>