Hypothesis 2: Gene L is necessary for phage propagation.
- Made new primer 4 and primer 4 T3485A mixes from the source at 100 and 10 μM.
- 50 μL WP-PCR reaction:
- 37 μL H2O
- 5 μL 10X reaction buffer
- 1.5 μL 3.2 nM ΦX174 template (~ 0.1 nM final)
- 0.5 μL 25 mM dNTPs mix
- 1 μL PfuUltra I DNA polymerase
- Aliquot 2 × 18 μL:
- 2 μL 10 μM primer 4 mix
- 2 μL 100 μM primer 4 mix
- Cycling parameters:
- 95 °C 2 m
- 95 °C 30 s
- 58° C 30 s
- 72 °C 15 m
- Repeat 2-4 an additional 29 times for 30 total cycles
- 72 °C 30 m
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