Team:Paris Saclay/Notebook/September/9

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Contents

Lab Work

By Mélanie

D-Lemon scent

extraction of TOPO plasmide with PS

Extraction with a Dna Kit extraction

[photo]

I failed

PCR

Transformation show some clone so I do a PCR.

component volume
H2O 41.5μl
buffer 5μl
dNTPs 1μl
Primer 1 1μl
Primer 2 1μl
bacteria (about 1µl = 1 colony)
Dream taq 0.5μl

Primer used:

Pu Pr (universal primer)


Cycle step Temperature Time Cycle

Bacteria lysis

95°C

5 min

1

Denaturation 94°C 30 s 25
Annealing 50°C 25 s 25
Extension 72°C 1 min 25
Final extension 72°C 10 min 1
Final extension 8°C hold 1

0909 PCR CLONE.jpg

0909 PCR CLONE LS.jpg

photo 1 GS (8 clones) ans cad (8 clones) photo 2 LS (8 clones)

After these results I choose some clones to do a liquid culture