M13 Experiments on the preparation of phage-derived vectors
M13 Experiments on the preparation of phage-derived vectors
M13 phage is a filamentous phage that does not lyse bacterial cells after infection, but only utilizes intracellular material to complete its own proliferation, and after assembling into complete viral particles, it is secreted by the host cell, which can still continue to grow and divide. It is an ideal plasmid carrier in molecular biology. Some nucleotide sequences in the spacer region will not affect the replication of M13DNA even if they are mutated or inserted into exogenous DNA fragments, which provides conditions for the construction of cloning vectors with M13DNA.
Operation method
carrier-derived method
Materials and Instruments
Plasmids Move 1. Fresh overnight culture buckwheat from a single colony was diluted at 1 : 50 in 2xYT culture medium (1~5 ml) containing appropriate antibiotics and incubated at 37°C until OD600=0.1. For more product details, please visit Aladdin Scientific website.
YT Medium
Centrifuge Spectrophotometer Shaker
2. Cells were infected at 1, 10, 20, 50 MOI and incubated at 37°C for 4.5 h under vigorous shaking.3. Centrifuge at 4 000 g for 10 min, collect the supernatant and inactivate by heating at 65℃ for 15 min.4. Extract single-stranded DNA from the supernatant and analyze the single-stranded DNA on an agarose gel with helper phage as a control.