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==
 +
===B - 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
 +
 +
 +
===D - lemon scent===
 +
 +
==== Electrophoresis of the 5 PCR of Limonene synthase (BBa762100) ====
 +
"by Hoang Vu"
 +
 +
(photo)
 +
 +
====Electrophoresis of the same 5 PCR pooled====
 +
''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.
 +
 +
(photo)
 +
 +
====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)
 +
 +
 +
{{Team:Paris_Saclay/notebook_footer}}
{{Team:Paris_Saclay/notebook_footer}}

Revision as of 14:38, 28 August 2014

Contents

Thursday 28th August

Lab Work

B - 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


D - lemon scent

Electrophoresis of the 5 PCR of Limonene synthase (BBa762100)

"by Hoang Vu"

(photo)

Electrophoresis of the same 5 PCR pooled

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.

(photo)

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)


Back to the calendar

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