Team:Paris Saclay/Notebook/August/18

From 2014.igem.org

(Difference between revisions)
(→D - Lemon Scent)
(→PCR of pCola)
Line 150: Line 150:
Five tubes were prepared. Protocol was modified to match the one used for pCola (PCR conducted on [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/8#PCR 8th August]).  
Five tubes were prepared. Protocol was modified to match the one used for pCola (PCR conducted on [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/8#PCR 8th August]).  
-
Primers used : iPS91bis and iPS82
+
{| class="wikitable centre" width="50%"
 +
|+ PCR of BBa_K762100
 +
|-
 +
! scope=col | components
 +
! scope=col | volumes
 +
|-
 +
|DNA of p cola
 +
|1μl
 +
|-
 +
|green GoTaq buffer 5X
 +
|10μl
 +
|-
 +
|GoTaq enzyme
 +
|0.5µl
 +
|-
 +
|H<sub>2</sub>O
 +
|31.25µl
 +
|-
 +
|MgCl<sub>2</sub>
 +
|4µl
 +
|-
 +
|dNTP
 +
|1µl
 +
|-
 +
|IPS82
 +
|1µl
 +
|-
 +
|IPS91bis
 +
|1µl
 +
|-
 +
|GoTaq polymerase
 +
|.25µl
 +
|}
====Electrophoresis of the 3 PCR====
====Electrophoresis of the 3 PCR====

Revision as of 10:05, 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
PCR cycle
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.

PCR of BBa_K517003
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).

PCR of BBa_K762100
components volumes
DNA of BBa_K517003 1μl
green GoTaq buffer 5X 10μl
GoTaq enzyme 0.5µl
H2O 31.25µl
MgCl2 4µl
dNTP 1µl
IPS66 1µl
IPS67 1µl
GoTaq polymerase .25µl

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).

PCR of BBa_K762100
components volumes
DNA of p cola 1μl
green GoTaq buffer 5X 10μl
GoTaq enzyme 0.5µl
H2O 31.25µl
MgCl2 4µl
dNTP 1µl
IPS82 1µl
IPS91bis 1µl
GoTaq polymerase .25µl

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

PCR of BBa_K517003
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

Back to the calendar

Retrieved from "http://2014.igem.org/Team:Paris_Saclay/Notebook/August/18"