Team:Paris Saclay/Notebook/August/8

From 2014.igem.org

(Difference between revisions)
(→Friday 8th August)
m (→Photo of the Day)
 
(13 intermediate revisions not shown)
Line 1: Line 1:
 +
{{Team:Paris_Saclay/notebook_header}}
=Friday 8th August=
=Friday 8th August=
==Lab work==
==Lab work==
-
===D - Lemon Scent===
+
===Salicylate Inducible Suppressing System===
 +
====Transformation of competent E.coli cells====
 +
''by Fabio''
-
We have done PCR with pcola (geranyl synthase) and BBa517003:
+
We transformed in DH5α the Ligation's result made [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/7#Ligation yesterday] ('''BBa_K1372000''' with '''BBa_B0015'''). ''The samples were put at 37°C for 10 hours, then we put them at room temperature until monday morning)''
-
Green GotaqBuffer = 10µl
+
'''Samples:'''
 +
# Sample 2 μl
 +
# Sample 1 μl
 +
# DH5α Control in LB
 +
# DH5α Control in LB + Cm
 +
'''Results on 11th August morning:'''
 +
# Success, grown very well (> 200 colonies)
 +
# Success, grown well (> 100 colonies)
 +
# Success, a film of bacteria has settled
 +
# Success, nothing has grown
 +
[https://2014.igem.org/Team:Paris_Saclay/Protocols/Transformation_of_supercompetent_Ecoli_cells_with_CaCl2 Protocol]
 +
===Lemon Scent===
 +
====PCR====
 +
''by Mélanie & Pierre''
 +
We have done a PCR with pcola (geranyl synthase) and BBa_K517003:
-
MgCl2 = 4µl  
+
* Green GotaqBuffer = 10µl
 +
* MgCl2 = 4µl  
 +
* dNTP  = 1µl
 +
* Primer A = 1µM
 +
* Primer B = 1µM
 +
* DNA = 1µl
 +
* Gotaq DNA polymerase = 0.25µl
-
dNTP  = 1µl
+
Oligo used : iPS81bis AND iPS82 (pcola) iPS69 and iPS68bis (BBa_K517003)
-
Primer A = 1µM
+
PCR cycles:
-
Primer B = 1µM
+
* 95° 2min
 +
* 95° 30sec
 +
* 49-55° 30 sec
 +
* 72° 2min
 +
* 71° 5min
-
DNA  = 1µl
+
====Electrophoresis====
 +
[[File:Paris Saclay-080814-Pierre-Mélanie-PCR.jpg|right|400px]]
 +
#10 µl of '''BBa_K517003'''
 +
#10 µl of '''BBa_K517003'''
 +
#10 µl of '''BBa_K517003'''
 +
#10 µl of '''BBa_K517003'''
 +
#10 µl of '''BBa_K517003'''
 +
#10 µl of '''p. cola'''
 +
#10 µl of '''p. cola'''
 +
#10 µl of '''p. cola'''
 +
#10 µl of '''p. cola'''
 +
#10 µl of '''p. cola'''
-
Gotaq DNA polymerase = 0.25µl
+
====Purification of the previous PCR====
 +
''by Pierre''
 +
From the 5 tubes of p.Cola I cleaned up two tubes.
 +
From the 5 tubes of BBa_K517003 I cleaned up two tubes.
-
Oligo used : iPS81bis AND iPS82 (pcola)      iPS69 and iPS68bis (BBa517003)
+
I did an electroforesis to check if the clening has worked. It did.
-
PCR cycle:
+
#1 µl of '''BBa_K517003'''
 +
#1 µl of '''p. cola'''
-
95° 2min
+
====Stock of pcola plasmides====
-
 
+
-
95° 30sec
+
-
 
+
-
49-55° 30 sec
+
-
 
+
-
72° 2min
+
-
 
+
-
71° 5min
+
-
 
+
-
 
+
-
====stock of pcola plasmide====
+
using the DNA kit extraction to extract pcola ()
using the DNA kit extraction to extract pcola ()
-
 
+
==Human Practices==
-
==Human Pratices==
+
===Art & Design===
===Art & Design===
''by Terry''
''by Terry''
Line 47: Line 77:
The mold is cut delicately and the test piece removed.
The mold is cut delicately and the test piece removed.
-
[[File:Photo agar test 003.jpg|950px]]
+
[[File:Photo agar test 003.jpg|750px|center]]
Then, the mold is placed back in the becher and filled up with regular agar ( 20mg/ml )
Then, the mold is placed back in the becher and filled up with regular agar ( 20mg/ml )
Line 58: Line 88:
Result : SUCCESS, I slightly remoistened the gel with water for an easier unmolding. The agar test piece is perfectly removed.
Result : SUCCESS, I slightly remoistened the gel with water for an easier unmolding. The agar test piece is perfectly removed.
-
[[File:Photo agar test 004.jpg|950px]]
+
[[File:Photo agar test 004.jpg|750px|center]]
 +
 
 +
==Photo of the Day==
 +
[[File:Paris Saclay 8_august.jpg|600px|center]]
 +
 
 +
 
 +
'''Members present:'''
 +
* Instructors and advisors: Alice, Solenne and Sylvie.
 +
* Students: Fabio, Hoang Vu, Eugène, Juliette, Mélanie, Pierre Romain, Sean and Terry
 +
 
 +
{{Team:Paris_Saclay/notebook_footer}}

Latest revision as of 14:49, 14 October 2014

Contents

Friday 8th August

Lab work

Salicylate Inducible Suppressing System

Transformation of competent E.coli cells

by Fabio

We transformed in DH5α the Ligation's result made yesterday (BBa_K1372000 with BBa_B0015). The samples were put at 37°C for 10 hours, then we put them at room temperature until monday morning)

Samples:

  1. Sample 2 μl
  2. Sample 1 μl
  3. DH5α Control in LB
  4. DH5α Control in LB + Cm

Results on 11th August morning:

  1. Success, grown very well (> 200 colonies)
  2. Success, grown well (> 100 colonies)
  3. Success, a film of bacteria has settled
  4. Success, nothing has grown

Protocol

Lemon Scent

PCR

by Mélanie & Pierre We have done a PCR with pcola (geranyl synthase) and BBa_K517003:

  • Green GotaqBuffer = 10µl
  • MgCl2 = 4µl
  • dNTP = 1µl
  • Primer A = 1µM
  • Primer B = 1µM
  • DNA = 1µl
  • Gotaq DNA polymerase = 0.25µl

Oligo used : iPS81bis AND iPS82 (pcola) iPS69 and iPS68bis (BBa_K517003)

PCR cycles:

  • 95° 2min
  • 95° 30sec
  • 49-55° 30 sec
  • 72° 2min
  • 71° 5min

Electrophoresis

Paris Saclay-080814-Pierre-Mélanie-PCR.jpg
  1. 10 µl of BBa_K517003
  2. 10 µl of BBa_K517003
  3. 10 µl of BBa_K517003
  4. 10 µl of BBa_K517003
  5. 10 µl of BBa_K517003
  6. 10 µl of p. cola
  7. 10 µl of p. cola
  8. 10 µl of p. cola
  9. 10 µl of p. cola
  10. 10 µl of p. cola

Purification of the previous PCR

by Pierre

From the 5 tubes of p.Cola I cleaned up two tubes. From the 5 tubes of BBa_K517003 I cleaned up two tubes.

I did an electroforesis to check if the clening has worked. It did.

  1. 1 µl of BBa_K517003
  2. 1 µl of p. cola

Stock of pcola plasmides

using the DNA kit extraction to extract pcola ()

Human Practices

Art & Design

by Terry

Sequel of yesterday molding attempt.

The mold is cut delicately and the test piece removed.

Photo agar test 003.jpg

Then, the mold is placed back in the becher and filled up with regular agar ( 20mg/ml )

Result : FAILURE, the agar test piece has been ripped when unmolding. It was stuck to the mold.

But the mold is still intact ! I cleaned it up and slightly covered the inward surface with liquid soap. The mold is filled up again and conserved in the refrigerator for 2 hours.

Result : SUCCESS, I slightly remoistened the gel with water for an easier unmolding. The agar test piece is perfectly removed.

Photo agar test 004.jpg

Photo of the Day

Paris Saclay 8 august.jpg


Members present:

  • Instructors and advisors: Alice, Solenne and Sylvie.
  • Students: Fabio, Hoang Vu, Eugène, Juliette, Mélanie, Pierre Romain, Sean and Terry

Back to the calendar

Retrieved from "http://2014.igem.org/Team:Paris_Saclay/Notebook/August/8"