I did antibiotic resistant gene transfer using mating experiment. The transconjugant was grown on antibiotic supplemented agar (antibiotics the donor and recipient resistant to). While the donor and recipient grow within 18-24 hours (in their respective antibiotic), the transconjugant only grow after 3-4 days in small number colonies. Is that mean the transconjugant have received the resistant gene or nothing changed if it takes too long to grow? The donor and recipient did not grow at all in the agar with combined antibiotics. I need to confirm this as I am going to do genome sequencing. Thanks
Hi there,
If the donor and recipient strains don't grow efficiently on the mixture of antibiotics then what is actually growing should be transformed cells. The observed delay in growth might be due to the time needed for the transferred resistance to express efficiently and to the dose of antibiotics used. Prior to global analysis by sequencing, I would go for a more rapid screen like PCR.
https://www.researchgate.net/post/How_can_we_distinguish_transconjugant_cells_from_recipient_cells_in_minimal_agar_medium
this may help you
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