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Confused ligation


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4 replies to this topic

#1 anonymous

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Posted 16 April 2001 - 09:00 PM

My amp plates are always flooded with satellite colonies even within a period of 16-18 hours of incubation. What do I do? Is there any other cheaper analog of carbencillin? I have tried spreading the amp also to counter heat inactivation while pouring- doesnt seem to help. help!

#2 anonymous

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Posted 16 April 2001 - 09:00 PM

i have same experience in the past, you can try to use a fresh make Amp plates i.e.use the plates which make in the same day.The problem will be less serious.

Good Luck!

#3 anonymous

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Posted 17 April 2001 - 09:00 PM

What's the Conc. you are using? You may use 100ug/ml. Another suggestion is to short the incubation time.

Good luck!


#4 anonymous

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Posted 23 April 2001 - 09:00 PM

Use lower insert to vector ratio (1:1 to 3:1) and lower DNA concentration. If protein is toxic you might need to use another vector.

#5 anonymous

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Posted 23 April 2001 - 09:00 PM

Clontech sells a PCR random mutagenesis kit.




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