5-fluoroorotic acid - (Sep/25/2001 )
u are looking for ura- cells. in a 5-FOA plate only ura- cells can grow. to look for these auxotrotrphs-first grow your transformant in rich media and then plate different dilutions on a rich plate. replica plate on to 5-FOA plates. colonies that appear after 2 days of incubation at 30o is what u are looking for.composition of 5-FOA platesyeast nitrogen base 6.7g, drop out mix-ura 2g, glucose 20g, uracil 50mg, 5-FOA 1g, agar 20g, d/water 1000ml,
All the bestmaitreyi
Please tell me method.I'm looking for a way of Yeast selection(transformant).
w303-1a(-Ura) transformed with pRS406(Selection maker is Ura3).Tihs transform is integrate type.Now, I Want to remove Ura3 maker(pop-out) using 5-FOA(5-fluoroorotic acid).But I don't know this method.Please tell me.