Hi All,
I have a very basic question to ask. We generally inoculate a single colony in small culture for overnight growth following transferring them into big conical and wait for the OD 600 to reach 0.6 to 0.8. Now, if you inoculate a single colony early in the morning(lets say 8 am) and by 8 pm to 9 pm you see the OD 600 is already 0.6, can we start our induction? Is it always necessary to do small inoculation first? I know this way is very convenient but I am just asking for the sake of argument. If I am growing a single colony to 0.6, the culture is still fresh right?
Hi! Growing fresh cultures is better because older cultures may have lost the plasmid, resulting in lower protein yield. I hope this helps!
The case you are explaining is not reproducible all the time, at least 1% inoculum is required for a culture to grow. This is the reason we usually prefer having a primary/smaller culture first, so that we add 1-2% of it to the secondary/larger culture. Inoculating a larger volume directly may give you growth but you can't be sure that this will happen every time, especially if the colony you are inoculating is coming from an old plate.
Hope this helps.
- Badges
- Science topic
- Similar topics
- Organizational Psychology
- Training