Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Animal and Molecular Model Systems > Yeast Forum
Register Search Today's Posts Mark Forums Read

Yeast Forum Discuss research on yeast the model organism. Post questions and topics on the molecular biology and genetics of Saccharomyces cerevisiae and other yeast species.


5'FOA selection

5'FOA selection - Yeast Forum

5'FOA selection - Discuss research on yeast the model organism. Post questions and topics on the molecular biology and genetics of Saccharomyces cerevisiae and other yeast species.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 05-25-2009, 06:29 PM
Pipette Filler
Points: 626, Level: 12 Points: 626, Level: 12 Points: 626, Level: 12
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: May 2009
Posts: 4
Thanks: 0
Thanked 0 Times in 0 Posts
Lightbulb 5'FOA selection



Hello,
I'm a first year grad student, and here is my latest quandry.

I am trying to knockout an integrated URA marker in my yeast line in order that I may use URA to select for the addition of another plasmid which I have prepared.
I have succesfully done this with one of my yeast lines, by plating onto a 5'FOA plate.

Any colonies that grew on the first 5'FOA plate were patched onto a second 5
'FOA plate simultaneously with a -URA plate.

Colonies that did grow on the second 5'FOA plate but not on the -URA plate were considered to NOT contain the URA marker anymore.

With the second yeast line that I am working on it has not been so easy. The yeast will commonly grow on the second generation 5'FOA plate AND the -URA plate, indicating the presence of some resistance to 5'FOA that is independent of the presence of URA marker.

In my most recent attempt, I noticed that the colonies on the second generation 5'FOA plate are all white, whereas the colonies on the URA plate are pink. Is there any signfiicance to this?

Any general tips about how YOU use 5'FOA for selection?

On advice of my mentor i have been transforming in a linear piece from the pRS316-URA containing plasmid to force recombination before plating on the first 5'FOA plate.

Thanks!
DJ
Reply With Quote
Reply

Tags
5foa , cerevisiae , selection , yeast


Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
Natural selection is proven wrong gim Biology Forum 3 05-15-2009 07:43 PM
Concentration of gentamycin required for the selection kp35@duke.edu Arabidopsis and Plant Biology 0 03-22-2009 05:25 PM
Blue White Cloning Selection Positive Colonies liono Molecular Cloning Forum 1 02-20-2008 09:45 AM
Gentamycin selection apurv Arabidopsis and Plant Biology 0 08-18-2006 12:23 AM
G418 selection Jayakumar, R Protocols and Methods Forum 0 11-17-2005 01:40 AM


All times are GMT. The time now is 12:32 AM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.11559 seconds with 16 queries