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The germinated embryos were transferred to a banana multiplication medium for generating multiple shoots of each transformation event (Fig. 2E, G and I).
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Putative transgenic plants, from many transformed cell colonies, were transferred back to the multiplication medium containing only Timentin for clonal micropropagation.
As a shoot multiplication medium which contains charcoal, LPch is generally used for between 28 42 days; growth on this medium would be dependent on the microorganisms present.
When shoots regenerated from a cell colony, they were labelled as a series for each transformed cell colony, and transferred to multiplication medium containing 200 mg·l-1 Timentin to suppress A. tumefaciens growth.
Plantlets formed from these explants were subcultured onto fresh medium for subsequent multiplication.
The best medium for adventitious bud multiplication was modified MS medium supplemented with 4.0 mg/L TDZ, 15% coconut water, 60 g/L sucrose and 0.1% activated charcoal (AC).
Water is not a favourable medium for the growth and multiplication of microorganisms, and yet many can survive in it long enough to carry infection to humans.
After 4 weeks, the regenerated shoots were transferred onto shoot elongation medium for elongation and bud multiplication.
The kanamycin-resistant calluses were then cultured on the fresh selection medium for further selection and multiplication.
After four weeks, the regenerated shoots were transferred into shoot elongation medium for elongation and bud multiplication Elongated shoots of about 2.5 cm were rooted on a rooting medium.
After this period, the calli were transferred to a shoot differentiation and protocorm - like body (PLB) multiplication and elongation medium (A10 medium) with A4 medium as control.
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