self ligation - condition (Mar/12/2008 )

HI I want to do a self ligation but I need to be in high dilution condition favoring intramolecular ligation.

so what should be the total volume for ligation let's day for 2ug DNA


thanks

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My understanding is that self-ligation is favoured if the DNA concentration is about 10 ng/ul (which means a 200 ul reaction for your 2ug of DNA!!!). Intermolecular ligation is favoured if the concentration is over 100 ng/ul (which is a 20 ul reaciton). You could split the difference and have a 100 ul reaction, or else try a range of concentrations to see which works best for you.

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