Team:Paris Saclay/Notebook/August/22

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(Ligation of LS/GS/PS in pPSI)
(Ligation of LS/GS/PS in pPSI)
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==== Ligation of LS/GS/PS in pPSI====
==== Ligation of LS/GS/PS in pPSI====
For each synthase, put:  
For each synthase, put:  
-
#10 µL of synthase
 
-
#2 µL of pPSI
 
-
#2 µL of buffer
 
-
#5 µL of water
 
-
#1 µL of ligase
 
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Revision as of 12:51, 22 August 2014

Contents

Friday 22nd August

Lab work

D - Lemon Scent

pPSI digestion

components volumes
pPSI 40μl
FastDigest green buffer 10X 5μl
PacI 2µl
H2O 2µl

pPSI dephosphorylation

After 2 hours, add 1.5µL of Alkaline Phosphatase (AP) and let it 1 hour.

Then, inactivation of AP at 65°C during 15 minutes.

We've made a electrophoresis to check the digestion.

(photo)

Ligation of LS/GS/PS in pPSI

For each synthase, put:

components volumes
synthase 10μl
pPSI 2μl
buffer 2µl
H2O 5µl
ligase 1µl

And let it in the freezer during 3 hours.


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