Team:Paris Saclay/Notebook/August/18

From 2014.igem.org

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(→D - Lemon Scent)
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We used the 3 previously PCR products to perform 3 reaction of ligation. The DNA insert used for each was : P6, L1 and G5  
We used the 3 previously PCR products to perform 3 reaction of ligation. The DNA insert used for each was : P6, L1 and G5  
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(2X)T4 ligase buffer 5μL
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{{Team:Paris_Saclay/notebook_footer}}
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{| class="wikitable centre" width="50%"
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pGEMT easy(50ng)   1μL
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|+ PCR of BBa_K517003
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|-
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DNA insert (PCR product) 3μL  
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! scope=col | components
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! scope=col | volumes
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Ligase 1μL
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|-
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|(2X)T4 ligase buffer
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vf : 10μL
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|5μL
 +
|-
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|pGEMT easy(50ng)  
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|1 μl
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|-
 +
|GoTaq enzyme
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|0.5 µl
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|-
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|DNA insert (GS or LS or PS PCR products)
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|3μL
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|-
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|Ligase
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|1 µl
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|}
16°C until afternoon and 4°C during the night
16°C until afternoon and 4°C during the night
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{{Team:Paris_Saclay/notebook_footer}}
 

Revision as of 15:06, 19 August 2014

Contents

Monday 18th August

Lab work

D - Lemon Scent

PCR of BBa_K517003

By Hoang Vu and Eugène

We realized a PCR to amplify the β-pinene synthase gene.

PCR of BBa_K517003
components volumes
DNA of GS 1 μl
green goTaq buffer 5X 10 μl
GoTaq enzyme 0.5 µl
H2O 29.5 µl
MgCl2 4 µl
dNTP 1µl
IPS68bis 2 µl
IPS69bis 2 µl

PCR of BBa_K762100

by Sean

Five tubes were prepared. Protocol was modified to match the one used for BBa_K517003 and p cola (PCR conducted on 8th August).

Primers used: iPS66 and iPS67.

PCR of pCola

by Laetitia and Melanie

We realized a PCR to amplify the geraniol synthase gene.

Five tubes were prepared. Protocol was modified to match the one used for pCola (PCR conducted on 8th August).

Primers used : iPS91bis and iPS82

Electrophoresis of the 3 PCR

(image)

Ligation of PCR products inside pGEMTeasy

by Laetitia

We used the 3 previously PCR products to perform 3 reaction of ligation. The DNA insert used for each was : P6, L1 and G5

Back to the calendar

PCR of BBa_K517003
components volumes
(2X)T4 ligase buffer 5μL
pGEMT easy(50ng) 1 μl
GoTaq enzyme 0.5 µl
DNA insert (GS or LS or PS PCR products) 3μL
Ligase 1 µl

16°C until afternoon and 4°C during the night

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