Algae, Tree, Herbs, Bush, Shrub, Grasses, Vines, Fern, Moss, Spermatophyta, Bryophyta, Fern Ally, Flower, Photosynthesis, Eukaryote, Prokaryote, carbohydrate, vitamins, amino acids, botany, lipids, proteins, cell, cell wall, biotechnology, metabolities, enzymes, agriculture, horticulture, agronomy, bryology, plaleobotany, phytochemistry, enthnobotany, anatomy, ecology, plant breeding, ecology, genetics, chlorophyll, chloroplast, gymnosperms, sporophytes, spores, seed, pollination, pollen, agriculture, horticulture, taxanomy, fungi, molecular biology, biochemistry, bioinfomatics, microbiology, fertilizers, insecticides, pesticides, herbicides, plant growth regulators, medicinal plants, herbal medicines, chemistry, cytogenetics, bryology, ethnobotany, plant pathology, methodolgy, research institutes, scientific journals, companies, farmer, scientists, plant nutrition
Select Language:
 
 
 
 
Main Menu
Please click the main subject to get the list of sub-categories
 
Services offered
 
 
 
 
  Section: Biotechnology Methods » Biochemistry
 
 
Please share with your friends:  
 
 

Estimation of DNA by the Diphenylamine Method

 
     
 

Principle
When DNA is treated with diphenylamine under acidic conditions, a blue compound is formed with the sharp observation maximum at 595 n. This reaction is produced by 2 deoxypentose in general and is not specific for DNA. In acid solution the straight from the deoxypentose is converted to the highly reactive b-hydroxy lerulin aldehyde, which reacts with diphenylamine to produce a blue complex. In DNA, only the deoxyribose of purine nucleotide reacts so that the value obtained represents one half of the total deoxyribose produced.

Materials

  • Commercial sample of DNA – 10 mg.
  • Buffer saline 0.15 mL/L NaCl, 0.15 mL/L sodium citrate made up to 500 mL.
  • Diphenyl amine reagent: dissolved in 10 gms of pure diphenyl amine in 1 liter of glacial acetic acid and add 25 mL of concentrated H2SO4. This solution must be prepared fresh.
  • Boiling water bath
 

 
Procedure
Dissolve 10 gms of nucleic acid in 50 mL of buffer saline. Remove 2 mL and add 4 mL of diphenylamine. Remove 2 mL and add 4 mL of DPA reagent. Heat on a boiling water bath for 10 nm. Cool and read the extension at 595 nm. Read the test and standard against nucleic acid and the commercial sample for DNA.

Result: Concentration of DNA present in the given sample ............... mg/2 mL.
 
     
 
 
     




     
 
Copyrights 2012 © Biocyclopedia.com | Disclaimer