Team:Paris Saclay/Notebook/August/22

From 2014.igem.org

(Difference between revisions)
(→Friday 22nd August)
(→D - Lemon Scent)
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==Lab work==
==Lab work==
===D - Lemon Scent===
===D - Lemon Scent===
-
====PpsI digestion====
+
====pPSI digestion====
40µl pPSI
40µl pPSI
Line 11: Line 11:
5µL FAST digest buffer
5µL FAST digest buffer
 +
 +
====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 chech the digestion.
 +
(photo)
 +
 +
==== Ligation of LS/GS/PS in pPSI====
 +
For each synthase,put:
 +
#10 µL of synthase
 +
#2 µL of pPSI
 +
#2 µL of buffer
 +
#5 µL of water
 +
#1 µL of ligase
 +
 +
And let it in the freezer during 3 hours.
 +
 +
{{Team:Paris_Saclay/notebook_footer}}
{{Team:Paris_Saclay/notebook_footer}}

Revision as of 10:11, 22 August 2014

Contents

Friday 22nd August

Lab work

D - Lemon Scent

pPSI digestion

40µl pPSI

2µl H2O

2µL PacI

5µL FAST digest buffer

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 chech 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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