Team:Paris Saclay/Notebook/August/22

From 2014.igem.org

(Difference between revisions)
(→pPSI digestion)
(→Ligation of LS/GS/PS in pPSI)
Line 41: Line 41:
#5 µL of water
#5 µL of water
#1 µL of ligase
#1 µL of ligase
 +
 +
{| class="wikitable centre" width="25%"
 +
|+ 
 +
|-
 +
! scope=col | components
 +
! scope=col | volumes
 +
|-
 +
|synthase
 +
|10μl
 +
|-
 +
|pPSI
 +
|2μl
 +
|-
 +
|buffer
 +
|2µl
 +
|-
 +
|H<sub>2</sub>O
 +
|5µl
 +
|-
 +
|ligase
 +
|1µl
 +
|}
And let it in the freezer during 3 hours.
And let it in the freezer during 3 hours.

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:

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

And let it in the freezer during 3 hours.


Back to the calendar

Retrieved from "http://2014.igem.org/Team:Paris_Saclay/Notebook/August/22"