Estimation of DNA by Diphenylamine Method
Автор: DBT VNSGU
Загружено: 2022-02-09
Просмотров: 5277
Описание:
Estimation of DNA in unknown sample using Diphenylamine Reagent
• Label 1 to 7 test tubes as Blank, 01, 02, 03, 04, 05 and Unknown sample.
• Pipette out different dilutions of Standard DNA solution (100µg/mL) in the test tubes.
• Make final volume as 2.0 mL using 0.5N Perchloric acid.
• In the test tube labelled as Blank, pipette out 2.0 mL of 0.5N Perchloric acid.
• Pipette out 2.0 mL of Unknown sample in the last test tube.
• Add 4.0 mL of Diphenylamine Reagent to all the tubes.
• Cover the tubes with Foil paper.
• Keep the tubes in boiling water bath for 15 minutes.
• Remove the tubes and let them cool.
• Record the absorbance at 600nm using Blank as Reagent blank.
• Draw a standard curve of Absorbance v/s DNA concentration.
• From the absorbance of Unknown sample, determine the amount of DNA in it.
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