Team:UNITN-Trento/Notebook/Labposts/06/40

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Latest revision as of 07:51, 3 October 2013

{ "date" : "2013-06-19", "author" : "gabriele-emil-bruno", "title" : "pSB1C3 (quite) an apocalipse and SAMsynthetase the (quite) revenge
linearization of pSB1C3 ad inocula of SAMsynthetase", "content" : "Today we (GG, ET, BA) tried to linearize pSB1C3 (with poor results), with this protocol, using the circular DNA samples of Tuesday (18-06): Emil 117.3ng/microl, Gabriele 184.1ng/microl.

Then we prepared a gel (1% GellyPhor gel) and checked for the products.

Gel electrophoresis of the products

Loaded samples	Result
1kb ladder	-
empty	-
B1	OK
B2	OK
B3	OK
E1	NO
E2	NO
E3	NO
G1	NO
G2	OK
G3	OK
empty	-
1kb ladder	-

Unfortunately, some of the PCR reactions did not succeed!

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.

Plate pSB1C3+R0010

Plate pSB1C3+SAMsynthetase ctrl

Plate pSB1C3+SAMsynthetase 1:1

Plate pSB1C3+SAMsynthetase 1:2

Plate pSB1C3+SAMsynthetase 1:4

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 (1µl of 34mg/ml Cloramphenycol stock solution for 1ml of LB).

In the afternoon Gabriele performed again the same PCR protocol to linearize pSB1C3. He loaded the E1-3 samples again, as double-check.

Gel electrophoresis of the products (Gabriele)

Loaded samples	Result
G1A	NO
G2A	OK
G3A	OK
G4A	OK
1kb ladder	-
E1	NO
E2	NO
E3	NO

", "tags" : "SAMsynthetase-pSB1C3" }

@@ Line 1: / Line 1: @@
 {
-	"date" : "2013-06-21",
+	"date" : "2013-06-19",
-	"author" : "thomas",
+	"author" : "gabriele-emil-bruno",
-	"title" : "Inocula of AraCpBAD + EFE in pSB1C3",
+	"title" : "pSB1C3 (quite) an apocalipse and SAMsynthetase the (quite) revenge<html><br/></html>''linearization of pSB1C3 ad inocula of SAMsynthetase''",
-	"content" : "I inoculated the previously transformed AraCpBAD + EFE in pSB1C3 into 5ml of LB containing Chloramphenicol.{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Inocula image|<html><center><img id=\"post_img\" src=\"https://static.igem.org/mediawiki/2013/2/2e/Tn-20130621-Inocula_TP.JPG\" width=\"400px\"/></center></html>}}As you can see, it seems that all the inocula grown. I will proceed with the purification and the screening test!",
+	"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>",
-	"tags" : "EFE-AraCpBAD"
+	"tags" : "SAMsynthetase-pSB1C3"
 }