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Agrobacterium cells were plated on solid AB medium containing antibiotic and incubated at 28°C for 3 days.
Most of the His+ clones (~ 83%) exhibited growth on medium containing antibiotic G418, indicating the stability of the transformants.
There is a possibility that plasmids may get lost from bacteria during infection and in such case, selecting bacteria on medium containing antibiotic will underestimate the bacterial load.
The cDNA-inserts of bacterial clones were PCR-amplified directly from aliquots of bacterial cultures grown overnight (LB medium, containing antibiotic selection).
Six hours later cell culture medium was removed and replaced with complete medium and after 48 hours the post-transfection medium was replaced with complete selection medium containing antibiotic G418 to a final concentration of 400 µg/ml.
Similar(55)
This overnight culture was inoculated into fresh LB medium containing antibiotics to an OD600 of 0.05 0.1 (1:50 dilution of the overnight culture).
The HAM was washed with DMEM (Dulbecco's modified eagle's medium) containing antibiotics under the laminar air flow to make it free from blood clots.
The cultures were inoculated in fresh LB medium containing antibiotics and incubated at 37°C until the optical density at 600 nm reached 0.8.
The testes were put in ice cold DMEM medium containing antibiotics and then carefully detunicated.
The homogenates were resuspended in 3 ml sterile brain heart infusion (BHI) medium containing antibiotics as described [51].
Bacterial cultures were grown overnight in LB medium containing antibiotics without agitation and washed in 0.9% NaCl before infection.
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