The stratagene bacterial match II two hydrid system uses M9 ammonium as base media for growth. The recipient strain has a hisB mutation which is complemented with yeast his3 gene upon protein-protein interaction allowing cells to grow in the presence of 3-AT.
The manufacture says to supplement M9 with adenine, thiamine and his drop out solution which has 17 amino acids excepct histidine, methionine, and cysteine.
I don´t see any sense for this supplementation as the recipient strain should be able to produce all aminoacids except his for ammonium provided in the M9 media. Has anyone tried to used regular M9 without His drop out solution?
Hi there,
You may use non supplemented M9 medium provided the recipient strain has no auxotrophy but growth rate will be much slower than on supplemented medium.
We use nitrogen base along with amino acid drop out in yeast two-hybrid assay. Having the nitrogen base in the medium make yeast to grow fast. So I guess supplementing M9 medium would speed up the growth of bacteria.
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