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SUMMARY:Improved Methods of Nucleic Acid Quantification via UV Spectrophoto
 metry
LOCATION:Chemistry A101
TZID:America/Denver
DTSTART:20191101T000000
UID:2026-04-25-00-11-13@natsci.colostate.edu
DTSTAMP:20260425T001113
Description:Literature Seminar\nThe most commonly used method of quantifica
 tion for single- and double-stranded nucleic acid species is UV spectropho
 tometry. Beer’s law relates the absorbance of a sample with the pathleng
 th\, concentration and attenuation coefficient (ε). For nucleic acids\, a
 n average mass attenuation coefficient is applied to approximately quantif
 y the species independent of base composition. There are two main issues t
 hat need to be considered when quantifying nucleic acids with spectrophoto
 metry: hypochromicity in double-stranded species\, and absorbance of conta
 minants at the incident wavelength. The hypochromicity phenomenon explains
  that double-stranded nucleic acids absorb less light than their correspon
 ding single-strands due to base stacking interactions. Additionally\, comp
 ounds like phenol are used in DNA/RNA extraction\, and a phenol contaminat
 ion can significantly change the absorbance at the incident 260 nm wavelen
 gth. As a means of improving the accuracy of quantification by UV spectrop
 hotometry\, Nwokeoji et al proposed a denaturing step in 50% DMSO to elimi
 nate the impact of hypochromicity on absorbance measurements. This techniq
 ue is especially useful for quantifying double-stranded RNA\, as tradition
 al methods of denaturing RNA can result in hydrolysis. This talk will focu
 s on previous work that seeks to quantify RNA and DNA as well as the newly
  proposed methods of RNA and DNA sample preparation that allow for a highe
 r level of accuracy to be achieved when quantifying with spectrophotometry
 . 4:00 pm
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