Team:Paris Saclay/Notebook/September/3

(Difference between revisions)

Revision as of 12:00, 7 September 2014

We made a classic exttraction of plasmids from the liquid cultures.

Check by electrophoresis if it worked and send it for sequencing.

by Mélanie

Electrophoresis of the PCR made Yesterday

well 2-3 = PCR with Dream taq or Vent taq

(well 4-5 = checking of the pps5 Extraction)

The PCR have success so I use the PCR clean up kit to purify it

Electrophoresis after purification

We already have some pPSI digested and dephosphorelated

So I do a ligation:

2 hours at room temperature and over night at 4°

Plasmid exctraction using the kit (picture is here)

digestion by SalI

Electrophoresis

We saw that we have something strang with our plasmid but we wil check it tomorrow with a good ladder (the ladder here was normally on the well 1)

We digest the plasmid by HindIII to direct our insert

well 1 = ladder well 2-3-4 = pPS3 Well 5-6-7 = pPS4

results are very strange

@@ Line 13: / Line 13: @@
 Electrophoresis of the PCR made [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/2#Limonene_synthase_PCR Yesterday]
-[photo]
-The PCR have success so I use the PCR clean up kit to purify it
-Electrophoresis after purification
 [[File:0309 PCR LS + extraction pPS5.jpg|400px]]
@@ Line 25: / Line 19: @@
 (well 4-5 = checking of the pps5 Extraction)
+The PCR have success so I use the PCR clean up kit to purify it
+Electrophoresis after purification
+[[File:0309 purif LS.jpg|400px]]
 ====Ligation====