Team:Paris Saclay/Notebook/July/29

From 2014.igem.org

(Difference between revisions)
(→Tuesday 29th July)
(→Tuesday 29th July)
Line 32: Line 32:
====Electrophoresis====
====Electrophoresis====
''by Romain''
''by Romain''
-
 
-
===E - Salicylate Inducible Suppressing System===
 
-
====Digestion====
 
-
''by Fabio''
 
-
 
-
BioBrick '''BBa_J61051''' (Salicylate promoter + NahR)
 
-
 
-
====Electrophoresis====
 
-
''by Fabio''
 
-
 
-
'''Results:'''
 
-
 
-
First process to verify the digestion
 
-
 
-
Second one to separate both parts (only with J61051).
 
-
 
-
'''Members there''':
 
-
* Instructors and advisors: Alice, Solenne and Sylvie.
 
-
* Students: Arnaud, Fabio, Romain, Sean and Terry.
 
===C - Lemon scent===
===C - Lemon scent===
Line 92: Line 73:
| 0.25μl
| 0.25μl
|}
|}
 +
 +
===E - Salicylate Inducible Suppressing System===
 +
====Digestion====
 +
''by Fabio''
 +
 +
BioBrick '''BBa_J61051''' (Salicylate promoter + NahR)
 +
 +
====Electrophoresis====
 +
''by Fabio''
 +
 +
'''Results:'''
 +
 +
First process to verify the digestion
 +
 +
Second one to separate both parts (only with J61051).
 +
 +
'''Members there''':
 +
* Instructors and advisors: Alice, Solenne and Sylvie.
 +
* Students: Arnaud, Fabio, Romain, Sean and Terry.
[https://2014.igem.org/Team:Paris_Saclay/Notebook Back to the calendar]
[https://2014.igem.org/Team:Paris_Saclay/Notebook Back to the calendar]

Revision as of 16:11, 29 July 2014

Contents

Tuesday 29th July

Lab Work

A - The frame

PCR

by Romain

StrainS used: E. coli MG1655Z1 and E. coli MG1655, and oligonucleotides used: FTR-Apra-F and FTR-Apra-R.

Protocol:

  • 218µl of H20 MilliQ
  • 80µl GoTaq buffer 5X
  • 16µl oligonucleotide FTR-Apra-F (10mM)
  • 16µl oligonucleotide FTR-Apra-R (10mM)
  • 12µl DMSO 10mM
  • 32µl MgCl2 (25mM)
  • 8µl dNTP (10mM)
  • 2µl of GoTaq enzyme
  • 2µl of different bacterial cultures in each tubes:
  1. Tube 1: MG1655Z1 10I
  2. Tube 2: MG1655 100II
  3. Tube 3: MG1655 50I
  4. Tube 4: MG1655 100I
  5. Tube 5: MG1655Z1 10II
  6. Tube 6: MG1655 50II
  7. Tube 7: MG1655Z1 10I positive control with plasmid

Electrophoresis

by Romain

C - Lemon scent

PCR of BBa_K762100

by Sean

Oligonucleotides used: iPS66, iPS67

Oligonucleotides were diluted twice from 100µM to 50µM.

Protocol

Add into a PCR tube the following:

Component For a total volume of 50μl
H2O 35.5μl
Phusion buffer 5X 10μl
dNTPs 1μl
iPS66 1μl
iPS67 1μl
BBa_K762100 1μl
Phusion DNA polymerase 0.25μl

E - Salicylate Inducible Suppressing System

Digestion

by Fabio

BioBrick BBa_J61051 (Salicylate promoter + NahR)

Electrophoresis

by Fabio

Results:

First process to verify the digestion

Second one to separate both parts (only with J61051).

Members there:

  • Instructors and advisors: Alice, Solenne and Sylvie.
  • Students: Arnaud, Fabio, Romain, Sean and Terry.

Back to the calendar

Retrieved from "http://2014.igem.org/Team:Paris_Saclay/Notebook/July/29"