Team:Paris Saclay/Notebook/July/29

From 2014.igem.org

(Difference between revisions)
(→Tuesday 29th July)
(→Lab Work)
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* Instructors and advisors: Alice, Solenne and Sylvie.
* Instructors and advisors: Alice, Solenne and Sylvie.
* Students: Arnaud, Fabio, Romain, Sean and Terry.
* Students: Arnaud, Fabio, Romain, Sean and Terry.
 +
 +
===C - Lemon scent===
 +
 +
====PCR of BBa_K762100====
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''by Sean''
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 +
Oligonucleotides used: iPS66, iPS67
 +
 +
Oligonucleotides were diluted twice from 100µM to 50µM.
 +
 +
'''Protocol'''
 +
 +
Add into a PCR tube the following:
 +
 +
{| class="wikitable centre" width="50%"
 +
|+
 +
|-
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! scope=col | Component
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! scope=col | Example for 50μl
 +
|-
 +
|H<sub>2</sub>O
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|35.5μl
 +
|-
 +
| Phusion buffer 5X
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| 10μl
 +
|-
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| dNTPs
 +
| 1μl
 +
|-
 +
| iPS66
 +
| 1μl
 +
|-
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| iPS67
 +
| 1μl
 +
|-
 +
| BBa_K762100
 +
| 1μl
 +
|-
 +
| Phusion DNA polymerase
 +
| 0.25μl
 +
|}
[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 15:51, 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 Fabio & Romain

Members there:

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

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 Example for 50μl
H2O 35.5μl
Phusion buffer 5X 10μl
dNTPs 1μl
iPS66 1μl
iPS67 1μl
BBa_K762100 1μl
Phusion DNA polymerase 0.25μl

Back to the calendar

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