Phage gDNA extraction (PCI)
Materials required:
For phage lysis
- Phage lysate: 500uL high titer, filtered with 0.22uM filters
- DNAse I: 10U total
- RNAse A: 1uL of 100mg/mL (0.1 mg total)
- DNAse buffer: 50uL of 10X
- EDTA: 20uL of 0.5M
- Proteinase K: 2.5U total
- SDS: 25uL of 10%
For DNA extraction
- TE-saturated Phenol:Choloroform:Isoamyl alcohol 25:24:1
- Chloroform:Isoamyl alcohol 24:1
Sources
https://www.pnas.org/doi/10.1073/pnas.2104592118#sec-3 At the Bench: A Laboratory Navigator (pg 284, pg 145)