k***g
发帖数: 4904 | 1 如果我想每天取得相似浓度的fresh culture，可不可以每天从同一浓度冻存的样品中
取出一些细菌，等比例稀释在media里incubate相同时间？这样能大致保持养出的细菌
浓度相近吗？ | I***a
发帖数: 13467 | | k***g
发帖数: 4904 | 3 俺现在就是每次都测OD，好麻烦啊
【在 I***a 的大作中提到】
: 不好，
: 你要计数或是测OD 才比较准一点，
| b******y
发帖数: 627 | 4 If you are doing protein expression, you don't need to measure OD600. I
always estimate by eye. That is because I never follow induction at OD600 =
0.6 rule. This rule is bullshit for a lot of, if not most, proteins. So far,
I have worked on more than 100 proteins (and their variants). None of them
require induction at OD600 = 0.6.
Normally I estimate the OD600 at 0.6~0.8. I reduce the temperature to 16~20
degree for an hour. Add 2 gram/liter of glucose and 1 mM IPTG. Don't worry
too much if you find your culture is pretty cloudy due to whatever reasons.
Chances are, you will have higher yield than otherwise.
However, I will be more cautious when I culture M9, especially in D2O. |
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