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Blunt-Cohesive ligation


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#1 laurequillo

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Posted 20 August 2010 - 01:23 AM

Hi guys!

One quick question. When you perform a ligation with both ends (Blunt and cohesive), do you add PEG to the reaction and do you increase the amount of ligase?

Thanks!
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#2 maral

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Posted 20 August 2010 - 01:37 AM

Hi,
i performed ligation with cohesive ends without using PEG.the enzyme was T4 DNA ligase. but my friend who performed ligation with blunt ends use PEG to improve final efficiancy.

#3 laurequillo

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Posted 20 August 2010 - 02:10 AM

View Postmaral, on 20 August 2010 - 01:37 AM, said:

Hi,
i performed ligation with cohesive ends without using PEG.the enzyme was T4 DNA ligase. but my friend who performed ligation with blunt ends use PEG to improve final efficiancy.

yeah sure, but I want to know when you have both ends how you do it. I have done them before without PEG and without add more ligase, I just want to know how you guys do it.

thanks
"He must be very ignorant for he answers every question he is asked" Voltaire

"This is SPARTA!"

"I´m the goddamn batman"




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