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Title: How to determine the concentration of plasmid DNA? Post by: florapt93 on Oct 23, 2012 You want to determine the concentration of plasmid DNA (i.e. it is double-stranded) in a stock tube. You dilute it by mixing 22 microlitres of your stock DNA with water to obtain a final volume of 1 mL. You obtain an absorbance reading at 260 nm of 0.15 using a standard 1-cm-wide cuvette. What is the concentration of DNA in your original stock tube? Title: How to determine the concentration of plasmid DNA? Post by: Nursmonica on Oct 23, 2012 double-stranded DNA = 50 ug x OD260 x dilution factor
http://matcmadison.edu/biotech/resources/methods/labManual/unit_4/exercise_15.htm good luck. Title: How to determine the concentration of plasmid DNA? Post by: onemustfall! on Oct 23, 2012 An OD260 of 1 is equivalent to 50 micrograms/ml.
Concentration of DNA was 0.15 X 50 micrograms/ml = 7.5 micrograms/ml. You diluted 22 microlitres into 1000 Your initial concentration was (1000/22) X 7.5 micrograms/ml = 340.91 micrograms/ml. |