Team:Paris Saclay/Notebook/August/28

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{{Team:Paris_Saclay/notebook_header}}
{{Team:Paris_Saclay/notebook_header}}
=Thursday 28th August=
=Thursday 28th August=
 +
 +
==Lab Work==
 +
===Construction of the fusion protein (color)===
 +
 +
====Transformation of DH5a by PSB1C3+chromoprotein====
 +
''by Mélanie''
 +
 +
--> a compléter
 +
 +
====Addition of adenines at the ends of PCR fragment of chromoprotein====
 +
''by Laetitia''
 +
 +
{| class="wikitable centre" width="50%"
 +
|+ 
 +
|-
 +
! scope=col | components
 +
! scope=col | volumes
 +
|-
 +
|H<sub>2</sub>O
 +
|1 μl
 +
|-
 +
|Gotaq buffer 5X
 +
|1µl
 +
|-
 +
|dATP 1mM
 +
|2 µl
 +
|-
 +
|Chromoprotein gene fragment (30ng/µL)
 +
|5µL
 +
|-
 +
|Gotaq polymerase
 +
|1µl
 +
|-
 +
|}
 +
 +
1h- 70°C
 +
 +
==== Ligation of the PCR fragment of chromoprotein + AAA with pGEMTeasy====
 +
''by Laetitia''
 +
 +
{| class="wikitable centre" width="50%"
 +
|+ 
 +
|-
 +
! scope=col | components
 +
! scope=col | volumes
 +
|-
 +
|2X ligation buffer T4 DNA ligase
 +
|10 μl
 +
|-
 +
|pGEMTeasy
 +
|1µl
 +
|-
 +
|Ligation mix
 +
|7 µl
 +
|-
 +
|Ligase
 +
|2µL
 +
|-
 +
|}
 +
 +
4h - 16°C
 +
 +
 +
===lemon scent===
 +
 +
==== Electrophoresis of 5 digestions of pPS5 by HindIII to check if the insert of CAD is in the right position  ====
 +
 +
[[File:280814 HV degisetion.jpg|300px|right]]
 +
''by Hoang Vu''
 +
 +
1-5: pPS5 (pJBEI+CAD) digested by HindIII
 +
 +
6: pPS1 (pJBEI+Apra), our witness
 +
 +
7-8: ladder
 +
 +
====Electrophoresis of the same 5 PCR pooled====
 +
 +
[[File:280814 Laetitia verif decoup.jpg|300px|left]]
 +
''by Laetitia''
 +
 +
Gel agarose 0,8% - 100V
 +
 +
The totality of the 5 PCR have been used. Then, we cut the lane corresponding of the Limonene synthase gene on a UV table.
 +
 +
 +
====Purification of the PCR product of Limonene synthase (BBa762100)====
 +
''by Laetitia''
 +
We used the kit PCR-Clean-UP to extract the DNA from the agarose gel. Elution volume : 15µl
 +
 +
 +
==Photo of the Day==
 +
[[File:Paris Saclay 28_august.jpg|400px|center]]
 +
 +
'''Members present:'''
 +
* Instructors and advisors: Alice, Solenne and Sylvie.
 +
* Students: Eugène, Hoang Vu, Laëtitia, Mélanie, Romain, Sean and Terry.
 +
{{Team:Paris_Saclay/notebook_footer}}
{{Team:Paris_Saclay/notebook_footer}}

Latest revision as of 15:28, 14 October 2014

Contents

Thursday 28th August

Lab Work

Construction of the fusion protein (color)

Transformation of DH5a by PSB1C3+chromoprotein

by Mélanie

--> a compléter

Addition of adenines at the ends of PCR fragment of chromoprotein

by Laetitia

components volumes
H2O 1 μl
Gotaq buffer 5X 1µl
dATP 1mM 2 µl
Chromoprotein gene fragment (30ng/µL) 5µL
Gotaq polymerase 1µl

1h- 70°C

Ligation of the PCR fragment of chromoprotein + AAA with pGEMTeasy

by Laetitia

components volumes
2X ligation buffer T4 DNA ligase 10 μl
pGEMTeasy 1µl
Ligation mix 7 µl
Ligase 2µL

4h - 16°C


lemon scent

Electrophoresis of 5 digestions of pPS5 by HindIII to check if the insert of CAD is in the right position

280814 HV degisetion.jpg

by Hoang Vu

1-5: pPS5 (pJBEI+CAD) digested by HindIII

6: pPS1 (pJBEI+Apra), our witness

7-8: ladder

Electrophoresis of the same 5 PCR pooled

280814 Laetitia verif decoup.jpg

by Laetitia

Gel agarose 0,8% - 100V

The totality of the 5 PCR have been used. Then, we cut the lane corresponding of the Limonene synthase gene on a UV table.


Purification of the PCR product of Limonene synthase (BBa762100)

by Laetitia We used the kit PCR-Clean-UP to extract the DNA from the agarose gel. Elution volume : 15µl


Photo of the Day

Paris Saclay 28 august.jpg

Members present:

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

Back to the calendar

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