Estimation of DNA by Spectroscopy Method

a EXPERIMENT NO 5 DATE: 14.11.2022 ESTIMATION OF DNA BY SPECTROPHOTOMETRI METHOD AIM TO essimate quantitatively DNA by spectrophotometric method MATERIALS REQUIRED A UV spectrophotometer converte Stervle distilled water or TE Buffer PROCEDURE Switch on UV spectrophotometer, adjust wavelength 260 nm put on UV at 'o' absorbance with Herile DNA destilled 2 Sel instrument or TE buffer depending on volubility of dilute 3. water absorbance of sample ifon is two ingh and take sample Read the with distilled water or TE buffer 4. Note the values at OD26044280nm on again 5. calculate OD260/00,200 Formula DNAconcernalism = ODaco ,Dilution facto 7 x50 purity natio = 1260/A280 18-2.0 - Numbic and 18 - Presence of protein our 120 Phenol/cnconform contamination

10 IA 260 of ds DNA = 50 Mg/ml 1A 260 of SS u Mg/ml RNA 40 IA 260 of 55 oligonucleoticle = 33/1g/ml RESULTS Average DNA concentration at 00260= 1812 Mg/m / Average purity ratio = 1.9867, which comes in the range 1.8-2.0 Hence the absorption is because of nuleic Jane and /