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yeast transforamtion


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#1 lewisyi

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Posted 22 June 2002 - 03:27 PM

yeast transformation by LiAc Method. When I did this, I followed the protocol in lab. I added DMSO before heatshock instead of adding before HS as the kit told me to do. Now two days after transformation no colony shows up. My questions are:1 what is DMSO supposed to do in transformation ? 2. Is the DMSO killed the yeasts? 3. how many days are the typical for colonies to show up? Thanks

#2 mkl

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Posted 27 June 2002 - 08:23 AM

hi,
first,DMSO supposed to help the transformation of yeast.
I was working with the 2 hybrid and I had the same problems. so what I did is the following:
1) I changed the amont of PEG (40%).
2) I add DMSO when I incubate (1h and 30min)
3) the most important is no wash the cells with TE 1x, left them in the same solution and plate (if the volume is big, centrifuge and take off half of it)
now, following this steps, I have colonies.
I hope that help you and if you have questions, you can email me (matouab@yahoo.fr)
I hope that you indersteand my english
good luck

#3 lewisyi

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Posted 27 June 2002 - 05:04 PM

thank you very much. i will try your way. i will let you know my result. tahnks again.




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