Team:Paris Saclay/Notebook/August/18
From 2014.igem.org
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Revision as of 09:52, 22 August 2014
Contents |
Monday 18th August
Lab work
B - Construction of the fusion protein (color)
PCR of the chromoprotein
by Mélanie
components | volumes |
---|---|
H2O | 27μl |
5X Phusion buffer | 10µl |
dNTP 10mM | 1µl |
iPS83 | 1µl |
iPS84 | 1µl |
DNA | .5µl |
DMSO | 1.5µl |
Phusion | .5µl |
step | temperature (°C) | time (s) |
---|---|---|
1 | 98 | 30 |
2 | 98 | 10 |
3 | 58 | 30 |
4 | 72 | 45 |
5 | 72 | 600 |
D - Lemon Scent
PCR of BBa_K517003
By Hoang Vu and Eugène
We realized a PCR to amplify the β-pinene synthase gene.
components | volumes |
---|---|
DNA of BBa_K517003 | 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 : PS-6, LS-1 and GS-5
components | volumes |
---|---|
(2X)T4 ligase buffer | 5μL |
pGEMT easy(50ng) | 1 μl |
DNA insert (GS or LS or PS PCR products) | 3μL |
Ligase | 1 µl |
16°C until afternoon and 4°C during the night