Team:Paris Saclay/Notebook/August/22

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(Difference between revisions)
(→pPSI dephosphorylation)
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===D - Lemon Scent===
===D - Lemon Scent===
====pPSI digestion====
====pPSI digestion====
-
40µl pPSI
 
-
2µl H2O
+
{| class="wikitable centre" width="50%"
-
 
+
|+ 
-
2µL PacI
+
|-
-
 
+
! scope=col | components
-
5µL FAST digest buffer
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! scope=col | volumes
 +
|-
 +
|pPSI
 +
|40μl
 +
|-
 +
|FastDigest green buffer 10X
 +
|5μl
 +
|-
 +
|PacI
 +
|2µl
 +
|-
 +
|H<sub>2</sub>O
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|2µl
 +
|}
====pPSI dephosphorylation====
====pPSI dephosphorylation====

Revision as of 12:43, 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:

  1. 10 µL of synthase
  2. 2 µL of pPSI
  3. 2 µL of buffer
  4. 5 µL of water
  5. 1 µL of ligase

And let it in the freezer during 3 hours.


Back to the calendar

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