MagicMedia™ E. coli Expression Medium SoluPouch™ - FAQs

Q: Can temperatures lower than 30&degC be used with MagicMedia even though the suggested incubation temperatures are 37&degC and 30&degC?

Yes, expression of proteins at 18&degC and 25&degC has been validated using the Dual Temperature protocol as follows. Day 1: Inoculate LB + antibiotic with an expression colony and incubate overnight at 30&degC, 300 rpm. Day 2: Inoculate pre-warmed (30&degC) MagicMedia + antibiotic at 1:40 with overnight seed culture. Incubate culture at 30&degC, 300 rpm for 6-7 hours or until OD600 reaches >6. Transfer culture to either 18&degC or 25&degC, 300 rpm and incubate for 24-36 hours. Day 3: Harvest and analyze results. Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center .

查看更多产品信息 MagicMedia™ E. coli Expression Medium SoluPouch™ - FAQs (K6802)

4 个常见问题解答

How does expression in MagicMedia affect the solubility of proteins when compared to standard LB and IPTG induction?

In most cases, the ratio of soluble to insoluble proteins is not much different when compared to LB and IPTG. However, since more total protein is typically obtained in MagicMedia, one also obtains more total soluble protein.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

How does MagicMedia compare to LB-IPTG with regards to expression of toxic proteins?

Toxic protein expression has not been investigated using MagicMedia. However, the Dual Temperature protocol provided in the manual can be tried. This may be a situation where LB/IPTG induction has an advantage since parameters (time, temperature, and IPTG concentration) are easily adjusted for optimal expression.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

Does MagicMedia have options to express and induce at mid-log phase?

No. Protein expression in MagicMedia occurs gradually over an 18-24 hour period, with detectable levels of expression beginning 6 hrs after a 1:40 seed inoculation with incubation at 30°C, 300 rpm.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

Can temperatures lower than 30°C be used with MagicMedia even though the suggested incubation temperatures are 37°C and 30°C?

Yes, expression of proteins at 18°C and 25°C has been validated using the Dual Temperature protocol as follows. Day 1: Inoculate LB + antibiotic with an expression colony and incubate overnight at 30°C, 300 rpm. Day 2: Inoculate pre-warmed (30°C) MagicMedia + antibiotic at 1:40 with overnight seed culture. Incubate culture at 30°C, 300 rpm for 6-7 hours or until OD600 reaches >6. Transfer culture to either 18°C or 25°C, 300 rpm and incubate for 24-36 hours. Day 3: Harvest and analyze results.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.