Team:Paris Saclay/Notebook/September/8

From 2014.igem.org

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(PCR with bacteria)
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[photo]
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We identify that we have success with to clone of PS.
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====Liquide culture of clones====
 +
The PS clone that reveal to be ok are cultivate in LB + AMP médium

Revision as of 15:46, 9 September 2014

Contents

Lab Work

D-Lemon Scent

Checking PCR

Electrophoresis of the PCR made Friday

[photo]

PCR with bacteria

From the petri dish (made here, I select some clone to do a PCR : clones 1-2-3 for cad clones 4-5 for PS clones 6 for LS

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:

For cloning in Topo vector = Pu Pr (universal primer)

For pPS2 = iPS 66/67

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

[photo]

We identify that we have success with to clone of PS.

Liquide culture of clones

The PS clone that reveal to be ok are cultivate in LB + AMP médium