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文献和实验Expression Protocol in M9 Minimal Media via T7 Promoter
The following protocol has been used successfully to 15N or 13C/15N label our proteins using our pET1120/BL21(DE3) expression system: Preparing M9 minimal media begins with preparing a 5x stock solution of M9 salts. Generally, M9 salts contain
Expression Protocol in M9 Minimal Media via T7 Promoter
with preparing a 5x stock solution of M9 salts. Generally, M9 salts contain a nitrogen source in the form of NH4 Cl. Since we want to add a labeled nitrogen source, our 5x salts are prepared minus NH4 Cl. Standard 5 X M9 Minimal Media salts minus nitrogen source
Preparing a Selenomethionyl Protein
1M IPTG M9 media (minimal media) 1 Liter 5x M9 media: (sterile filtered) 30g Na2 HPO4 or 64g Na2 HPO4 -7H2 O 15g KH2 PO4 5g NH4 Cl 2.5g NaCl Dilute and autoclave before use. Amino acid
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