I'm working on Peanut nodulation gene knockout using pYLCRISIPR/Cas9... One set using culture media (co-culture) and (selection media supplemented with hygromycin B) since the Binary plasmid has hygromycin R gene. This is used to select the positive hairy root transformants.

The second set using soil media, the seedlings were infected with A. rhizogene carrying the targeted gene (5days old seedlings). Now the seedlings have grown and I think I can see some hairy roots in them. How do I select the positive hairy root transformants using hygromycin B solution in the soil media? I will really appreciate if anybody has done this before and want to shine some light on it.

Also for the inoculation of rhizobia to the positive hairy root transformant seedlings, how should I go about applying rhizobia both for the culture media seedlings and the soil media seedlings. ?

Thank you.

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