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Do you any information regardingYeast 5-FOA selection. - (Oct/16/2001 )

Do you any information regarding Yeast 5-FOA selection.

I was constructed Yeast integrating plasmid pRS406-ORF-Flag tag(Tag sequence was added C terminus).This plasmid was transformed. And transformant were selectedwith -Ura plete.I want to 'pop out' the ura maker by homologous recombination. Thereby, few days ago, I submit this forum.I was teached Outline and performed.But, I don't obtain colony that chromosomal gene was tagged at the extreme C terminus by integrating a pRS06(popped out Ura makar).

Please tell me in detail the 5-FOA selection.For example Screening, Clture etc...

-Gentaro Yasuda-

are u using controls?? why dont u try using a control when u do the selection so that u can be sure of the media and other conditions u are using. also i suggest u read some old papers on 5FOA selection. a medline search will tell u what to read. as far as your problem is concerned i cannot follow exactly what problem are u facing now. please do not froget that the frequency of getting a POP OUT is quite low so u may have to screen a significantly large no of colonies to get what u want.all the bestmaitreyi