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I dissolved the DNA in 1XTAE instead of TE buffer, will it affect ligation?

DNA ligation transformation 1xTAE buffer TE buffer

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#1 babethepig

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Posted 03 October 2013 - 08:41 AM

After dephosphorylating the vector, I extracted it with phenol/chloroform extraction and ethanol precipitation. However, in the last step after rinsing the DNA with 70% ethanol and drying the DNA, I accidentally dissolved the DNA with 1XTAE buffer instead of TE buffer. Now I want to use this DNA solution for ligation. Will it work? I am just adding 1ul of this vector solution to a 10ul ligation reaction 



#2 jerryshelly1

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Posted 03 October 2013 - 09:28 AM

You should be fine.  I once used a sigma kit and they gave you an alternative ligation buffer if you suspended your DNA in TAE. When you are finished with your ligation, I would consider purifying your product somehow. TAE has been shown to significantly decrease the transformation efficiency of E. coli.

 

Google search pulled up the relevant kit with alternative buffer:

http://www.sigmaaldr...tmp/lig2bul.pdf







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