IGEM:Imperial/2010/XylE team - Revision history

Madeline Rounds: /* Friday, 20-Aug-2010 */

2010-10-21T08:42:59Z

Friday, 20-Aug-2010

←Older revision		Revision as of 08:42, 21 October 2010
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	==Friday, 20-Aug-2010==		==Friday, 20-Aug-2010==
-	~~[[Image:J23101.jpg\|thumb\|200px\|The J23101 gene in a biobrick vector containg RFP gene]]~~	+
	* we run a gel on XylEGFP1 PCR reaction. Results: GFP was extended successfully, XylE extension <font color = red>'''FAILED'''</font> (too much non-specific annealing)		* we run a gel on XylEGFP1 PCR reaction. Results: GFP was extended successfully, XylE extension <font color = red>'''FAILED'''</font> (too much non-specific annealing)
	*catechol assay on 2hrs bench ligation of promoter,xylE and vector failed.		*catechol assay on 2hrs bench ligation of promoter,xylE and vector failed.

Madeline Rounds: /* Tues 12th Oct */

2010-10-16T14:07:57Z

Tues 12th Oct

←Older revision		Revision as of 14:07, 16 October 2010
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	*Transformations worked! we now have PVEG-XylE in 3k3 and reverse-xylE with lacI promoter (C-TEV vector)i did replica plates, catechol assay plates and overnights for minis. Will mini prep tmro and transform them into TOP10 and we should be good to go with all constructs.		*Transformations worked! we now have PVEG-XylE in 3k3 and reverse-xylE with lacI promoter (C-TEV vector)i did replica plates, catechol assay plates and overnights for minis. Will mini prep tmro and transform them into TOP10 and we should be good to go with all constructs.
	* Assays messed up, my fault :( 3k3 didnt respond to catechol, so i think that I picked a background colony, i wont pick it again. The results are a bit ridiculous and will probably be chucked.		* Assays messed up, my fault :( 3k3 didnt respond to catechol, so i think that I picked a background colony, i wont pick it again. The results are a bit ridiculous and will probably be chucked.
		+
		+	==Weds 13th Oct==
		+	mini prepped 3k3-Pveg-XylE-B0014 and went to the school workshop! set off overnight midi of culture 9
		+
		+	==Thurs 14th Oct==
		+	Midi prepped 3k3-Pveg
		+
		+	==Fri 15th==
		+	Transformed all my constructs into T0P10 3k3-J23101/pveg and PSB1C3-J23101/PVEG

	==Output Photo Gallery==		==Output Photo Gallery==

Madeline Rounds: /* Monday 11th Oct */

2010-10-12T17:14:52Z

Monday 11th Oct

←Older revision		Revision as of 17:14, 12 October 2010
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	*Doing data analysis with my last assay results comparing high copy (1C3) vs low copy (3K3)and the effects on XylE activity..boring		*Doing data analysis with my last assay results comparing high copy (1C3) vs low copy (3K3)and the effects on XylE activity..boring
	*Finalising the polo-shirt logo design, it's looking fiiiine, we're gonna look goooood ;)		*Finalising the polo-shirt logo design, it's looking fiiiine, we're gonna look goooood ;)
		+
		+	==Tues 12th Oct==
		+	*Transformations worked! we now have PVEG-XylE in 3k3 and reverse-xylE with lacI promoter (C-TEV vector)i did replica plates, catechol assay plates and overnights for minis. Will mini prep tmro and transform them into TOP10 and we should be good to go with all constructs.
		+	* Assays messed up, my fault :( 3k3 didnt respond to catechol, so i think that I picked a background colony, i wont pick it again. The results are a bit ridiculous and will probably be chucked.

	==Output Photo Gallery==		==Output Photo Gallery==

Madeline Rounds: /* Monday 11th Oct */

2010-10-11T14:09:35Z

Monday 11th Oct

←Older revision		Revision as of 14:09, 11 October 2010
Line 1,020:		Line 1,020:
	*Gonna perform assays with them this afternoon after some growing time :)))))))		*Gonna perform assays with them this afternoon after some growing time :)))))))
	*Doing data analysis with my last assay results comparing high copy (1C3) vs low copy (3K3)and the effects on XylE activity..boring		*Doing data analysis with my last assay results comparing high copy (1C3) vs low copy (3K3)and the effects on XylE activity..boring
		+	*Finalising the polo-shirt logo design, it's looking fiiiine, we're gonna look goooood ;)

	==Output Photo Gallery==		==Output Photo Gallery==

Madeline Rounds: /* Friday 8th Oct */

2010-10-11T14:08:51Z

Friday 8th Oct

←Older revision		Revision as of 14:08, 11 October 2010
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	*Chris is purifying digested PVeg-XylE-B0014-C3 to get the insert out and this will then be ran on a gel alongside cut (E+X) 3k3 to determine a ratio for ligation.		*Chris is purifying digested PVeg-XylE-B0014-C3 to get the insert out and this will then be ran on a gel alongside cut (E+X) 3k3 to determine a ratio for ligation.
	*Meeting today at 4pm, no one is here!! it'll be me florian ben and piotr attending..yikes		*Meeting today at 4pm, no one is here!! it'll be me florian ben and piotr attending..yikes
		+
		+	==Sunday 10th of the 10th of the 2010!!!!! and im in LAB ==
		+	*I did ligations of PVeg-XylE into 3k3 and of Reverse-XylE into the digested C-Tev LacI vector.
		+	*I also set up assay cultures for monday 2x PVegXE 2x XE-C3 2x XE-3K3 and 2x GFP-XylE!!!
		+
		+	==Monday 11th Oct==
		+	*No Nicolas Kylilis, what a douche!
		+	*Diluted my overnights 250ul into 5ml to grow up to around 0.5 OD
		+	*Gonna perform assays with them this afternoon after some growing time :)))))))
		+	*Doing data analysis with my last assay results comparing high copy (1C3) vs low copy (3K3)and the effects on XylE activity..boring

	==Output Photo Gallery==		==Output Photo Gallery==

Madeline Rounds: /* Thursday 7th Oct */

2010-10-08T14:19:02Z

Thursday 7th Oct

←Older revision		Revision as of 14:19, 8 October 2010
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	*Midi prepped PVEG-XylE-B0014 in PSB1C3; will digest out the insert (E+S) and put it into 3K3(E+X). Then both the midi (in PSB1C3) and the ligation 3k3-PVEG-XylE-B0014 can be transformed into TOP10. J23101-XylE-B0014 in 3K3 Vector also needs to be put into TOP10 currently only the PSB1C3 version is in TOP10. We need the rest for comparison testing as TOP10 is the more widely used testing strain.		*Midi prepped PVEG-XylE-B0014 in PSB1C3; will digest out the insert (E+S) and put it into 3K3(E+X). Then both the midi (in PSB1C3) and the ligation 3k3-PVEG-XylE-B0014 can be transformed into TOP10. J23101-XylE-B0014 in 3K3 Vector also needs to be put into TOP10 currently only the PSB1C3 version is in TOP10. We need the rest for comparison testing as TOP10 is the more widely used testing strain.
		+	*Florian kindly digested PVeg-XylE-B0014-C3 with E+S
		+	*Ran PCR to get out blunt PVeg-XylE-B0014 using primers. Did 3 reactions at variations around 60 degrees.
		+
		+	==Friday 8th Oct==
		+	Ran gel analysis of my PCRs 1, 2 and 3 appear to have worked. Samples 1 and 3* show stronger bands and so should be used in the next ligation step into the SPEC vector (which kyasha will be doing) then transformation into Bacillus (meeee.)
		+	*Chris is purifying digested PVeg-XylE-B0014-C3 to get the insert out and this will then be ran on a gel alongside cut (E+X) 3k3 to determine a ratio for ligation.
		+	*Meeting today at 4pm, no one is here!! it'll be me florian ben and piotr attending..yikes

	==Output Photo Gallery==		==Output Photo Gallery==

Madeline Rounds: /* Thursday 7th Oct */

2010-10-07T15:51:51Z

Thursday 7th Oct

←Older revision		Revision as of 15:51, 7 October 2010
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	*Niiiiiiiiiiiiiiick!!! you left the country! you were too ugly. good luck babe!		*Niiiiiiiiiiiiiiick!!! you left the country! you were too ugly. good luck babe!
		+
		+	*Midi prepped PVEG-XylE-B0014 in PSB1C3; will digest out the insert (E+S) and put it into 3K3(E+X). Then both the midi (in PSB1C3) and the ligation 3k3-PVEG-XylE-B0014 can be transformed into TOP10. J23101-XylE-B0014 in 3K3 Vector also needs to be put into TOP10 currently only the PSB1C3 version is in TOP10. We need the rest for comparison testing as TOP10 is the more widely used testing strain.

	==Output Photo Gallery==		==Output Photo Gallery==

Piotr D. Faba: /* Thursday 7th Oct */

2010-10-07T15:30:23Z

Thursday 7th Oct

←Older revision		Revision as of 15:30, 7 October 2010
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	==Thursday 7th Oct==		==Thursday 7th Oct==
	*Labpartner, goodmorning and from greece!! :p		*Labpartner, goodmorning and from greece!! :p
		+
		+	*Niiiiiiiiiiiiiiick!!! you left the country! you were too ugly. good luck babe!

	==Output Photo Gallery==		==Output Photo Gallery==

Nicolas Kylilis: /* Weds 6th Oct */

2010-10-07T11:05:23Z

Weds 6th Oct

←Older revision		Revision as of 11:05, 7 October 2010
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	*Just realised Im working with background colony 16!! disregard this after the gel run!		*Just realised Im working with background colony 16!! disregard this after the gel run!
	*I'm currently keeping my undigested PVeg-XylE minis in florians DNA box (orange lids and labels)		*I'm currently keeping my undigested PVeg-XylE minis in florians DNA box (orange lids and labels)
		+
		+
		+	==Thursday 7th Oct==
		+	*Labpartner, goodmorning and from greece!! :p

	==Output Photo Gallery==		==Output Photo Gallery==

Madeline Rounds: /* Weds 6th Oct */

2010-10-06T10:10:11Z

Weds 6th Oct

←Older revision		Revision as of 10:10, 6 October 2010
Line 994:		Line 994:
	*Will perform the same assays already performed by Nick with the 3k3-XylE constructs to compare results.		*Will perform the same assays already performed by Nick with the 3k3-XylE constructs to compare results.
	*Just realised Im working with background colony 16!! disregard this after the gel run!		*Just realised Im working with background colony 16!! disregard this after the gel run!
		+	*I'm currently keeping my undigested PVeg-XylE minis in florians DNA box (orange lids and labels)

	==Output Photo Gallery==		==Output Photo Gallery==