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Latest revision as of 14:53, 14 October 2014

Monday 18th August

Lab work

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

PS : pinene synthase

LS : limonen synthase

GS : geraniol synthase

PS1 - PS2 - PS3 - PS4 - PS5 - PS6 - LS1 - LS2 - LS3 - LS4 - LS5 - GS1 - GS2 - GS3 - GS4 - GS5 - L

We have PCR product !

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

Ligation of BBa_K517003
components	volumes
(2X)T4 ligase buffer	5μL
pGEMT easy(50ng)	1 μl
DNA insert : BBa_K517003 digestion product	3μL
Ligase	1 µl

Ligation of BBa_K762100
components	volumes
(2X)T4 ligase buffer	5μL
pGEMT easy(50ng)	1 μl
DNA insert : BBa_K762100 digestion product	3μL
Ligase	1 µl

Ligation of pCola
components	volumes
(2X)T4 ligase buffer	5μL
pGEMT easy(50ng)	1 μl
DNA insert : pCola digestion product	3μL
Ligase	1 µl

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

Human Practices

Art & Design

by Terry

Incubation of a preculture of E. Coli with plasmid FNR RBS AmylCP (coding for a blue chromoprotein) in our new M63 medium for 24h.

Photo of the Day

Members present:

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

Back to the calendar

@@ Line 2: / Line 2: @@
 =Monday 18th August=
 ==Lab work==
-===B - Construction of the fusion protein (color)===
+===Construction of the fusion protein (color)===
 ====PCR of the chromoprotein====
 ''by Mélanie''
@@ Line 185: / Line 185: @@
 ====Electrophoresis of the 3 PCR====
-(image)
+PS : pinene synthase
+LS : limonen synthase
+GS : geraniol synthase
+PS1 - PS2 - PS3 - PS4 - PS5 - PS6 - LS1 - LS2 - LS3 - LS4 - LS5 - GS1 - GS2 - GS3 - GS4 - GS5 - L
+[[File:Photo 1808.jpg|300px]]
+We have PCR product !
 ====Ligation of PCR products inside pGEMTeasy====
@@ Line 257: / Line 269: @@
 Incubation of a preculture of ''E. Coli'' with plasmid FNR RBS AmylCP (coding for a blue chromoprotein) in our new M63 medium for 24h.
+==Photo of the Day==
+[[File:Paris Saclay 18_august.jpg|600px|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/August/18