Team:Paris/19 August 2009

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Latest revision as of 12:06, 10 October 2009

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NoteBook

June
M	T	W	T	F	S	S
[http://2009.igem.org/Team:Paris/1_June_2009 1]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/2_June_2009&action=edit 2]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/3_June_2009&action=edit 3]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/4_June_2009&action=edit 4]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/5_June_2009&action=edit 5]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/6_June_2009&action=edit 6]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/7_June_2009&action=edit 7]
[http://2009.igem.org/wiki/index.php?title=Team:Paris/8_June_2009&action=edit 8]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/9_June_2009&action=edit 9]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/10_June_2009&action=edit 10]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/11_June_2009&action=edit 11]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/12_June_2009&action=edit 12]	[http://2009.igem.org/Team:Paris/13_June_2009 13]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/14_June_2009&action=edit 14]
[http://2009.igem.org/wiki/index.php?title=Team:Paris/15_June_2009&action=edit 15]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/16_June_2009&action=edit 16]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/17_June_2009&action=edit 17]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/18_June_2009&action=edit 18]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/19_June_2009&action=edit 19]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/20_June_2009&action=edit 20]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/21_June_2009&action=edit 21]
[http://2009.igem.org/wiki/index.php?title=Team:Paris/22_June_2009&action=edit 22]	[http://2009.igem.org/Team:Paris/23_June_2009 23]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/24_June_2009&action=edit 24]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/25_June_2009&action=edit 25]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/26_June_2009&action=edit 26]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/27_June_2009&action=edit 27]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/28_June_2009&action=edit 28]
[http://2009.igem.org/wiki/index.php?title=Team:Paris/29_June_2009&action=edit 29]	[http://2009.igem.org/Team:Paris/30_June_2009 30]

July
M	T	W	T	F	S	S
		[http://2009.igem.org/wiki/index.php?title=Team:Paris/1_July_2009&action=edit 1]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/2_July_2009&action=edit 2]	[http://2009.igem.org/Team:Paris/3_July_2009 3]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/4_July_2009&action=edit 4]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/5_July_2009&action=edit 5]
[http://2009.igem.org/wiki/index.php?title=Team:Paris/6_July_2009&action=edit 6]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/7_July_2009&action=edit 7]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/8_July_2009&action=edit 8]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/9_July_2009&action=edit 9]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/10_July_2009&action=edit 10]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/11_July_2009&action=edit 11]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/12_July_2009&action=edit 12]
[http://2009.igem.org/Team:Paris/13_July_2009 13]	[http://2009.igem.org/Team:Paris/14_July_2009 14]	[http://2009.igem.org/Team:Paris/15_July_2009 15]	[http://2009.igem.org/Team:Paris/16_July_2009 16]	[http://2009.igem.org/Team:Paris/17_July_2009 17]	[http://2009.igem.org/Team:Paris/18_July_2009 18]	[http://2009.igem.org/Team:Paris/19_July_2009 19]
[http://2009.igem.org/Team:Paris/20_July_2009 20]	[http://2009.igem.org/Team:Paris/21_July_2009 21]	[http://2009.igem.org/Team:Paris/22_July_2009 22]	[http://2009.igem.org/Team:Paris/23_July_2009 23]	[http://2009.igem.org/Team:Paris/24_July_2009 24]	[http://2009.igem.org/Team:Paris/25_July_2009 25]	[http://2009.igem.org/Team:Paris/26_July_2009 26]
[http://2009.igem.org/Team:Paris/27_July_2009 27]	[http://2009.igem.org/Team:Paris/28_July_2009 28]	[http://2009.igem.org/Team:Paris/29_July_2009 29]	[http://2009.igem.org/Team:Paris/30_July_2009 30]	[http://2009.igem.org/Team:Paris/31_July_2009 31]

August
M	T	W	T	F	S	S
					[http://2009.igem.org/Team:Paris/1_August_2009 1]	[http://2009.igem.org/Team:Paris/2_August_2009 2]
[http://2009.igem.org/Team:Paris/3_August_2009 3]	[http://2009.igem.org/Team:Paris/4_August_2009 4]	[http://2009.igem.org/Team:Paris/5_August_2009 5]	[http://2009.igem.org/Team:Paris/6_August_2009 6]	[http://2009.igem.org/Team:Paris/7_August_2009 7]	[http://2009.igem.org/Team:Paris/8_August_2009 8]	[http://2009.igem.org/Team:Paris/9_August_2009 9]
[http://2009.igem.org/Team:Paris/10_August_2009 10]	[http://2009.igem.org/Team:Paris/11_August_2009 11]	[http://2009.igem.org/Team:Paris/12_August_2009 12]	[http://2009.igem.org/Team:Paris/13_August_2009 13]	[http://2009.igem.org/Team:Paris/14_August_2009 14]	[http://2009.igem.org/Team:Paris/15_August_2009 15]	[http://2009.igem.org/Team:Paris/16_August_2009 16]
[http://2009.igem.org/Team:Paris/17_August_2009 17]	[http://2009.igem.org/Team:Paris/18_August_2009 18]	[http://2009.igem.org/Team:Paris/19_August_2009 19]	[http://2009.igem.org/Team:Paris/20_August_2009 20]	[http://2009.igem.org/Team:Paris/21_August_2009 21]	[http://2009.igem.org/Team:Paris/22_August_2009 22]	[http://2009.igem.org/Team:Paris/23_August_2009 23]
[http://2009.igem.org/Team:Paris/24_August_2009 24]	[http://2009.igem.org/Team:Paris/25_August_2009 25]	[http://2009.igem.org/Team:Paris/26_August_2009 26]	[http://2009.igem.org/Team:Paris/27_August_2009 27]	[http://2009.igem.org/Team:Paris/28_August_2009 28]	[http://2009.igem.org/Team:Paris/29_August_2009 29]	[http://2009.igem.org/Team:Paris/30_August_2009 30]
[http://2009.igem.org/Team:Paris/31_August_2009 31]

September
M	T	W	T	F	S	S
	[http://2009.igem.org/Team:Paris/1_September_2009 1]	[http://2009.igem.org/Team:Paris/2_September_2009 2]	[http://2009.igem.org/Team:Paris/3_September_2009 3]	[http://2009.igem.org/Team:Paris/4_September_2009 4]	[http://2009.igem.org/Team:Paris/5_September_2009 5]	[http://2009.igem.org/Team:Paris/6_September_2009 6]
[http://2009.igem.org/Team:Paris/7_September_2009 7]	[http://2009.igem.org/Team:Paris/8_September_2009 8]	[http://2009.igem.org/Team:Paris/9_September_2009 9]	[http://2009.igem.org/Team:Paris/10_September_2009 10]	[http://2009.igem.org/Team:Paris/11_September_2009 11]	[http://2009.igem.org/Team:Paris/12_September_2009 12]	[http://2009.igem.org/Team:Paris/13_September_2009 13]
[http://2009.igem.org/Team:Paris/14_September_2009 14]	[http://2009.igem.org/Team:Paris/15_September_2009 15]	[http://2009.igem.org/Team:Paris/16_September_2009 16]	[http://2009.igem.org/Team:Paris/17_September_2009 17]	[http://2009.igem.org/Team:Paris/18_September_2009 18]	[http://2009.igem.org/Team:Paris/19_September_2009 19]	[http://2009.igem.org/Team:Paris/20_September_2009 20]
[http://2009.igem.org/Team:Paris/21_September_2009 21]	[http://2009.igem.org/Team:Paris/22_September_2009 22]	[http://2009.igem.org/Team:Paris/23_September_2009 23]	[http://2009.igem.org/Team:Paris/24_September_2009 24]	[http://2009.igem.org/Team:Paris/25_September_2009 25]	[http://2009.igem.org/Team:Paris/26_September_2009 26]	[http://2009.igem.org/Team:Paris/27_September_2009 27]
[http://2009.igem.org/Team:Paris/28_September_2009 28]	[http://2009.igem.org/Team:Paris/29_September_2009 29]	[http://2009.igem.org/Team:Paris/30_September_2009 30]

October
M	T	W	T	F	S	S
			[http://2009.igem.org/Team:Paris/1_October_2009 1]	[http://2009.igem.org/Team:Paris/2_October_2009 2]	[http://2009.igem.org/Team:Paris/3_October_2009 3]	[http://2009.igem.org/Team:Paris/4_October_2009 4]
[http://2009.igem.org/Team:Paris/5_October_2009 5]	[http://2009.igem.org/Team:Paris/6_October_2009 6]	[http://2009.igem.org/Team:Paris/7_October_2009 7]	[http://2009.igem.org/Team:Paris/8_October_2009 8]	[http://2009.igem.org/Team:Paris/9_October_2009 9]	[http://2009.igem.org/Team:Paris/10_October_2009 10]	[http://2009.igem.org/Team:Paris/11_October_2009 11]
[http://2009.igem.org/Team:Paris/12_October_2009 12]	[http://2009.igem.org/Team:Paris/13_October_2009 13]	[http://2009.igem.org/Team:Paris/14_October_2009 14]	[http://2009.igem.org/Team:Paris/15_October_2009 15]	[http://2009.igem.org/Team:Paris/16_October_2009 16]	[http://2009.igem.org/Team:Paris/17_October_2009 17]	[http://2009.igem.org/Team:Paris/18_October_2009 18]
[http://2009.igem.org/Team:Paris/19_October_2009 19]	[http://2009.igem.org/Team:Paris/20_October_2009 20]	[http://2009.igem.org/Team:Paris/21_October_2009 21]	[http://2009.igem.org/Team:Paris/22_October_2009 22]	[http://2009.igem.org/Team:Paris/23_October_2009 23]	[http://2009.igem.org/Team:Paris/24_October_2009 24]	[http://2009.igem.org/Team:Paris/25_October_2009 25]
[http://2009.igem.org/Team:Paris/26_October_2009 26]	[http://2009.igem.org/Team:Paris/27_October_2009 27]	[http://2009.igem.org/Team:Paris/28_October_2009 28]	[http://2009.igem.org/Team:Paris/29_October_2009 29]	[http://2009.igem.org/Team:Paris/30_October_2009 30]	[http://2009.igem.org/Team:Paris/31_October_2009 31]

← Yesterday - Tomorrow →

August 19th

Lab work

Sequencing w/ Bobode

5 clones pLac/RBS-TetR have to be sequenced

Clone 1, 2, 6, 7, 8 used (because of their high DNA concentration)

Transformation w/ Bobode

pSB1A3-TolRII ligation was transformed into P10 chemical competent cells, using the "heat shock" protocol

Glycerol stock w/ Charlie

Glycerol stock of :

S39 - pINTE3 (Col E3, AmpR)

S40 - pINTg3p (g3p Term, AmpR)

S41 - pINK2(TolRII, AmpR)

S42 - pTA (pTet-Col A, AmpR)

A28 A29

A28 OmpA signal (ex A11)

A29 FecR mutant

PCR 2 steps : 72°C 30s elongation

Gel verification Ok (OmpAsignal: weak)

Purification Ok

OD : A28=0,82 A29=0,92

Miniprep send to sequencing.

Miniprep DO with miniprep diluted 1/100:

ClyA colonie 3:1,40

ClyA colonie 7:1,60

ClyA colonie 8:0,8

ClyA colonie 10:1,65

The results were transformed into µg/mL

Mix for sequencing:

	Cly A colonie 3	Cly A colonie 7	Cly A colonie 8	Cly A colonie 10
Miniprep	4,29µL	3,75µL	7,50µL	3,64µL
Oligos	2,45µL	2,45µL	2,45µL	2,45µL
H2O	8,26µL	8,80µL	5,05µL	8,91µL

RFP PCR

RFP PCR using PSB1A3 and:

-O57+O58==>A30=RFP Nter

-O59+058==>A31=RFP

-O59+O60==>A33=RFP Cter

Mix:

H2O:32,5µL

Buffer 5x:10µL

dNTP:1µL

OF(1/10):2,5µL

OR(1/10):2,5µL

Matrice(PSB1A3 1/10):1µL

Phusiong:0,5µL

PCR Check

/100pb/A30/A31/A33/1kb

Digestion

Digestion PSB1A3 in XP

← Yesterday - Tomorrow →

@@ Line 4: / Line 4: @@
 ==NoteBook==
-{| style="color:#1b2c8a;background-color:#0;" cellpadding="0" cellspacing="1" border="0" bordercolor="#fff" width="100%" align="left"
+{{Paris2009_Calendar}}
-!align="center"|{{ #calendar: title=Team:Paris |year=2009 | month=06 }}
+{{Paris2009_Calendar_Link|18_August_2009|20_August_2009}}
-!align="center"|{{ #calendar: title=Team:Paris |year=2009 | month=07 }}
+<center> '''August 19th''' </center>
-!align="center"|{{ #calendar: title=Team:Paris |year=2009 | month=08 }}
-!align="center"|{{ #calendar: title=Team:Paris |year=2009 | month=09 }}
-!align="center"|{{ #calendar: title=Team:Paris |year=2009 | month=10 }}
-|}
 <html>
@@ Line 18: / Line 14: @@
 <div id="paris_content">
 </html>
-==Brain work==
-edit please ^^
-<html>
-</div>
-<div id="paris_content_boxtop">
-</div>
-<div id="paris_content">
-</html>
 ==Lab work==
@@ Line 69: / Line 50: @@
 <u>'''''Glycerol stock of ''''':</u>
-- pINTE3 (Col E3, ''AmpR'')
+S39 - pINTE3 (Col E3, ''AmpR'')
-- pTA (pTet-Col A, ''AmpR'')
+S40 - pINTg3p (g3p Term, ''AmpR'')
-- pINTg3p (g3p Term, ''AmpR'')
+S41 - pINK2(TolRII, ''AmpR'')
-- pINK2(TolRII, ''AmpR'')
+S42 - pTA (pTet-Col A, ''AmpR'')
 </div>
+<div class="caroline">A28 A29 </div>
+<div class="experience">
+A28 OmpA signal (ex A11)
+A29 FecR mutant
+PCR 2 steps : 72°C 30s elongation
+Gel verification Ok (OmpAsignal: weak)
-<html>
+Purification Ok
+OD : A28=0,82  A29=0,92
 </div>
-<div id="paris_content_boxtop">
+<div class="vicard">
+Miniprep send to sequencing.
 </div>
-<div id="paris_content">
+<div class="experience">
-</html>
-==To do list==
+Miniprep DO with miniprep diluted 1/100:
-edit please ^^
+ClyA colonie 3:1,40
+ClyA colonie 7:1,60
+ClyA colonie 8:0,8
+ClyA colonie 10:1,65
+The results were transformed into µg/mL
+Mix for sequencing:
+{|
+|- style="background: #b3d0E4; text-align: center; color:black; "
+|width=10%|
+|width=10%| Cly A colonie 3
+|width=10%| Cly A colonie 7
+|width=10%| Cly A colonie 8
+|width=10%| Cly A colonie 10
+|- style="background: #E0E9EF; text-align: center;"
+|width=10%| Miniprep
+|width=10%| 4,29µL
+|width=10%| 3,75µL
+|width=10%| 7,50µL
+|width=10%| 3,64µL
+|- style="background: #E0E9EF; text-align: center;"
+|width=10%| Oligos
+|width=10%| 2,45µL
+|width=10%| 2,45µL
+|width=10%| 2,45µL
+|width=10%| 2,45µL
+|- style="background: #E0E9EF; text-align: center;"
+|width=10%| H2O
+|width=10%| 8,26µL
+|width=10%| 8,80µL
+|width=10%| 5,05µL
+|width=10%| 8,91µL
+|}
+</div>
+<div class="vicard">
+RFP PCR
+</div>
+<div class="experience">
+RFP PCR using PSB1A3 and:
+-O57+O58==>A30=RFP Nter
+-O59+058==>A31=RFP
+-O59+O60==>A33=RFP Cter
+Mix:
+H2O:32,5µL
+Buffer 5x:10µL
+dNTP:1µL
+OF(1/10):2,5µL
+OR(1/10):2,5µL
+Matrice(PSB1A3 1/10):1µL
+Phusiong:0,5µL
+'''PCR Check'''
+/100pb/A30/A31/A33/1kb
+</div>
+<div class="vicard">
+Digestion
+</div>
+<div class="experience">
+Digestion PSB1A3 in XP
+</div>
+{{Paris2009_Calendar_Link|18_August_2009|20_August_2009}}