Team:UNITN-Trento/Notebook/Labposts/06/38

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Latest revision as of 07:50, 3 October 2013

{ "date" : "2013-06-19", "author" : "fabio-bruno", "title" : "bacillus subtilis promoters and screening (part 4)!!", "content" : " today we performed the screening protocol in order to verify the real presence of the promoters in the miniprep ( K823000a, K143012b) . To do that, we digested 600 ng of both templates with Ecor1-HF and Pst1-HF", "tags" : "Pveg-PliaG-PlepA" }

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 {
 	"date" : "2013-06-19",
-	"author" : "gabriele-emil-bruno",
+	"author" : "fabio-bruno",
-	"title" : "pSB1C3 (quite) an apocalipse and SAMsynthetase the (quite) revenge<html><br/></html>''linearization of pSB1C3 ad inocula of SAMsynthetase''",
+	"title" : "bacillus subtilis promoters and screening (part 4)!!",
-	"content" : "<html>Today we (GG, ET, BA) tried to linearize pSB1C3 (with poor results), with <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#pSB1C3-linearization-PCR\">this protocol</a>, using the circular DNA samples of Tuesday (18-06): Emil 117.3ng/microl, Gabriele 184.1ng/microl.<br /><br/>Then we prepared a gel (1% GellyPhor gel) and checked for the products.</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel electrophoresis of the products|<html><center><table class=\"tn-sp-table\"><tr><th>Loaded samples</th><th>Result</th></tr><tr><td>1kb ladder</td><td>-</td></tr><tr><td><i>empty</i></td><td>-</td></tr><tr><td>B1</td><td>OK</td></tr><tr><td>B2</td><td>OK</td></tr><tr><td>B3</td><td>OK</td></tr><tr><td>E1</td><td>NO</td></tr><tr><td>E2</td><td>NO</td></tr><tr><td>E3</td><td>NO</td></tr><tr><td>G1</td><td>NO</td></tr><tr><td>G2</td><td>OK</td></tr><tr><td>G3</td><td>OK</td></tr><tr><td><i>empty</i></td><td>-</td></tr><tr><td>1kb ladder</td><td>-</td></tr></table><img src=\"https://static.igem.org/mediawiki/2013/1/16/Tn-20130619-am_GG_AB_ET_pSB1C3_linearization_PCR_gel.jpg\" width=\"450px\" /></center></html>}}<html>Unfortunately, some of the PCR reactions did not succeed!<br/><br/>Then we (GG, ET) have proceeded with the inoculation of pSB1C3+SAMsynthetase (1:1, 1:2, 1:4) and of part R0010 (pLac). Unfortunately there were a few colonies also in the control.<br/></html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Plate pSB1C3+R0010|<html><center><img src=\"https://static.igem.org/mediawiki/2013/6/6b/Tn-20130619-am_ET_GG_pSB1C3_R0010.JPG\" height=\"450px\" /></center></html>}}<html></html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Plate pSB1C3+SAMsynthetase ctrl|<html><center><img src=\"https://static.igem.org/mediawiki/2013/6/63/Tn-20130619-am_ET_GG_pSB1C3_SAM_ctrl.jpg\" height=\"450px\" /></center></html>}}<html></html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Plate pSB1C3+SAMsynthetase 1:1|<html><center><img src=\"https://static.igem.org/mediawiki/2013/f/f2/Tn-20130619-am_ET_GG_pSB1C3_SAM_1to1.JPG\" height=\"450px\" /></center></html>}}<html></html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Plate pSB1C3+SAMsynthetase 1:2|<html><center><img src=\"https://static.igem.org/mediawiki/2013/0/0b/Tn-20130619-am_ET_GG_pSB1C3_SAM_1to2.JPG\" height=\"450px\" /></center></html>}}<html></html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Plate pSB1C3+SAMsynthetase 1:4|<html><center><img src=\"https://static.igem.org/mediawiki/2013/1/1a/Tn-20130619-am_ET_GG_pSB1C3_SAM_1to4.JPG\" height=\"450px\" /></center></html>}}<html>The inocula were performed in plastic culture tubes (15ml) instead of the glass ones, because we do not know how to sterilize the latter. The tubes were filled with 5ml of LB with Cloramphenycol (1&micro;l of 34mg/ml Cloramphenycol stock solution for 1ml of LB).<br/><br/>In the afternoon Gabriele performed again the same <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#pSB1C3-linearization-PCR\">PCR protocol</a> to linearize pSB1C3. He loaded the E1-3 samples again, as double-check.</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel electrophoresis of the products (Gabriele)|<html><center><table class=\"tn-sp-table\"><tr><th>Loaded samples</th><th>Result</th></tr><tr><td>G1A</td><td>NO</td></tr><tr><td>G2A</td><td>OK</td></tr><tr><td>G3A</td><td>OK</td></tr><tr><td>G4A</td><td>OK</td></tr><tr><td>1kb ladder</td><td>-</td></tr><tr><td>E1</td><td>NO</td></tr><tr><td>E2</td><td>NO</td></tr><tr><td>E3</td><td>NO</td></tr></table><img src=\"https://static.igem.org/mediawiki/2013/c/c0/Tn-20130619-pm_GG_pSB1C3_linearization.jpg\" width=\"450px\" /></center></html>}}<html></html>",
+	"content" : "<html>  today we performed the screening protocol in order to verify the real presence of the promoters in the miniprep ( K823000a, K143012b) . To do that, we digested 600 ng of both templates with Ecor1-HF and Pst1-HF</html>",
-	"tags" : "SAMsynthetase-pSB1C3"
+	"tags" : "Pveg-PliaG-PlepA"
 }