Team:Paris Bettencourt/Notebook/Phage Sensor/Monday 23rd September.html

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     <td><b>Thermocycler Protocol: NEB Quick-Load</b></td>
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     <td><b>Thermocycler Protocol: NEB Phusion</b></td>
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Revision as of 12:46, 2 October 2013

Detect

Monday 23rd September

Gels and PCR

Gel
100V, 1%, 1,5h

Picture: 13/09/23_SPCR16_17_corresponding primers

PCR
SPCR 16 (normal phusion at 48°C)

Reagent Volume
x1
Nuclease-free water 37.25 µl
5x Phusion HF Buffer 10 µl
10 mM dNTPs 1 µl
Forward Primer (10 uM) 0.5 µl
Reverse Primer (10 uM) 0.5 µl
Template Plasmid 0.25 µl
Phusion DNA Polymerase 0.5 µl
Total Volume 50 µl



Thermocycler Protocol: NEB Phusion
Temp Time
Start 98°C 30 sec Melt
Cycle 1 98°C 5 sec Melt 35 cycles
Cycle 2 72°C 30 sec Anneal
Cycle 3 72°C 30 sec per kb Extend
Finish 72 °C 5 min Extand
Store 10°C Forever Store



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