Team:Paris Saclay/Notebook/August/22
From 2014.igem.org
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===D - Lemon Scent=== | ===D - Lemon Scent=== | ||
====pPSI digestion==== | ====pPSI digestion==== | ||
- | |||
- | + | {| class="wikitable centre" width="50%" | |
- | + | |+ | |
- | + | |- | |
- | + | ! scope=col | components | |
- | + | ! scope=col | volumes | |
+ | |- | ||
+ | |pPSI | ||
+ | |40μl | ||
+ | |- | ||
+ | |FastDigest green buffer 10X | ||
+ | |5μl | ||
+ | |- | ||
+ | |PacI | ||
+ | |2µl | ||
+ | |- | ||
+ | |H<sub>2</sub>O | ||
+ | |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:
- 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.