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

Detect

ASDF

Monday 23^rd September

Gels and PCR

Gel
100V, 1%, 1,5h

Picture: 13/09/23_SPCR16_17_corresponding primers - PCRs didn't worked


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