Hello everyone!
I need to grow bacteria for maxiprep from a glycerol stock of TOP10 E. coli. Usually I prepare a starter culture of glycerol stock for 8 h and after that I dilute this and grow a maxiprep. But, is possible to grow bacteria for maxiprep directly from glycerol stock without a previous step of starter culture?
Thanks!
In principle the problem is performance, you must start a crop and go through all phases of growth: adaptation, logarithmic or exponential growth, stationary phase and cell death. The logical condition is to recover your bacteria in the logarithmic growth phase, which will depend on fast-growing or slow-growing bacteria, waiting for hours, they are fast-growing bacteria. When you preserve a culture in glycerol and refrigerate it at -70 degrees centigrade, you stop that growth, because you have a 50% mixture of bacteria in some culture medium and glycerol, in sterile conditions.
assuming that there is no strong selection against your plasmid because of over expression, then it should work fine. This is not optimal, I prefer to streak and pick a single colony, but you certainly can grow and prep directly from a glycerol stock.
Yes but not advised! The health of your culture and the amount of lyzed biomass are critical factor to obtain a good amount of pure plasmid. My advice to you: forget about the DIY kit option! Send us your plasmids as we can prep them for less than it takes you to do it. We typically obtain better results than you would with a kit in terms of DNA quantity and quality. See for yourself at https://www.biozilla.bio/plasmid-dna-purification
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology
- Microorganisms
- Bacteria