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

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Detect

Monday 23^rd 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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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