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cloning- ligation - (Nov/29/2013 )


I am a bit mixed up. This combination is for ligation procedure in cloning . I was wondering ng means how much DNA or vector in microliter in the mixture? 


 25ng Vector DNA 

 75ng Insert DNA 
 Ligase Buffer (1μL/10μL reaction for 10X buffer, and 2μL/10μL reaction for 5X buffer)
 0.5-1μL T4 DNA Ligase 
 H20 to a total of 10μL


Thank you very much for your time.


25 nanogram/microliter for insert DNA

75 nanogram/microliter for vector DNA


According to your recipe, you should use 25 ng of vector for the ligation and 75 ng for the insert. The amount of µL foe each will depend on the concentration of insert and vector. For instance, if your vector is at 50 ng/µL, you should use 0.5 µL in order to put 25 ng in the ligation mixture.


Hope it helps :)


Thank you very much "Enthusiast", I got it smile.png .