作者Ren (wisc)
看板Biotech
标题Re: [问题] 想问大家induction时所选择的OD600值
时间Sun Sep 16 00:11:38 2007
※ 引述《mournermind (阿蒙)》之铭言：
: 我们实验室长久以来的方法大概就是
: 1. single colony -> 养12hr以上 OD600约1.XX
: 2. 以约1/10的菌液加入新鲜LB中 OD600约0.1 进行refresh
: 3. 约2~3hr後 OD600 约0.5进行IPTG induce
: 4. 取定点时间induced的菌看蛋白表现
: 可是我在pET mannual中所看到的方法大概是
: 1. single colony -> 养到 OD600约 0.5
: 2. 以3ml的菌液加入100ml新鲜LB中进行refresh
: 3. 约2~3hr後 OD600 约0.5 ~ 1 进行IPTG induce
: 4. 取定点时间induced的菌看蛋白表现
补充一下：
1. Single colony -> 2~5mL. OD any, but must be turbid. (it's then
usually >0D 0.5.)
2. spin down if the next media is not LB. -> transfer to 20~ 50mL LB.
3. Typically after 8hrs, it will
OVER OD 1.0
4. Transfer into 1L LB. Carefully moniter the OD. ~OD=1.0 to
induce. Usually, it should be ok to induce at around OD 1.0~1.2.
5. Doubling time should not be more than 75min,
in my system.
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※ 编辑: Ren 来自: 76.201.144.184 (09/16 00:13)