Team:Paris/19 August 2009

From 2009.igem.org

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==NoteBook==
==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>
<html>
Line 19: Line 15:
</html>
</html>
 +
==Lab work==
 +
<div class="charlotte">
 +
Sequencing w/ Bobode
 +
</div>
 +
<div class="experience">
-
==Brain work==
 
 +
5 clones pLac/RBS-TetR have to be sequenced
-
edit please ^^
+
Clone 1, 2, 6, 7, 8 used (because of their high DNA concentration)
-
<html>
 
</div>
</div>
-
<div id="paris_content_boxtop">
 
-
</div>
 
-
<div id="paris_content">
 
-
</html>
 
-
==Lab work==
 
<div class="charlotte">
<div class="charlotte">
-
Sequencing w/ Bobode
+
Transformation w/ Bobode
</div>
</div>
<div class="experience">
<div class="experience">
-
5 clones pLac/RBS-TetR have to be sequenced
 
-
Clone 1, 2, 6, 7, 8 used (be cause of their high DNA concentration)
+
pSB1A3-TolRII ligation was transformed into P10 chemical competent cells, using the "heat shock" protocol
Line 48: Line 42:
-
<div class="charlotte">
+
<div class="romain">
-
Transformation w/ Bobode
+
Glycerol stock w/ Charlie
</div>
</div>
<div class="experience">
<div class="experience">
-
pSB1A3-TolRII ligation was transformed into P10 chemical competent cells, using the "heat shock" protocol
+
 
 +
<u>'''''Glycerol stock of ''''':</u>
 +
 
 +
S39 - pINTE3 (Col E3, ''AmpR'')
 +
 
 +
S40 - pINTg3p (g3p Term, ''AmpR'')
 +
 
 +
S41 - pINK2(TolRII, ''AmpR'')
 +
 
 +
S42 - pTA (pTet-Col A, ''AmpR'')
</div>
</div>
 +
<div class="caroline">A28 A29 </div>
 +
<div class="experience">
 +
A28 OmpA signal (ex A11)
-
<html>
+
A29 FecR mutant
 +
 
 +
PCR 2 steps : 72°C 30s elongation
 +
 
 +
Gel verification Ok (OmpAsignal: weak)
 +
 
 +
Purification Ok
 +
 
 +
OD : A28=0,82  A29=0,92
</div>
</div>
-
<div id="paris_content_boxtop">
+
 
 +
<div class="vicard">
 +
Miniprep send to sequencing.
</div>
</div>
-
<div id="paris_content">
+
<div class="experience">
-
</html>
+
 +
Miniprep DO with miniprep diluted 1/100:
 +
ClyA colonie 3:1,40
-
<div class="romain">
+
ClyA colonie 7:1,60
-
Glycerol stock w/ Charlie
+
 
 +
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>
<div class="experience">
<div class="experience">
-
Glycerol stock of :
+
RFP PCR using PSB1A3 and:
-
- pINTE3 (Col E3, ''AmpR'')
+
-O57+O58==>A30=RFP Nter
-
- pTA (pTet-Col A, ''AmpR'')
+
-O59+058==>A31=RFP
-
- pINTg3p (g3p Term, ''AmpR'')
+
-O59+O60==>A33=RFP Cter
-
- pINK2(TolRII, ''AmpR'')
+
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>
-
==To do list==
+
<div class="vicard">
 +
Digestion
 +
</div>
 +
<div class="experience">
-
edit please ^^
+
Digestion PSB1A3 in XP
 +
 
 +
</div>
 +
{{Paris2009_Calendar_Link|18_August_2009|20_August_2009}}

Latest revision as of 12:06, 10 October 2009

NoteBook

June
MTWTFSS
[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
MTWTFSS
    [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
MTWTFSS
          [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
MTWTFSS
  [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
MTWTFSS
      [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 →

Retrieved from "http://2009.igem.org/Team:Paris/19_August_2009"