Lab 1 SPECTROPHOTOMETRIC ANALYSIS OF NUCLEIC ACIDS

Lab 1 SPECTROPHOTOMETRIC ANALYSIS OF NUCLEIC ACIDS - Office...

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Office hours: Tuesday 4-5 ZHS 360A BioSpace
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Quiz Q1 : At which wavelength does DNA have the maximum absorbance? (2pts) Q2: Which two properties of a DNA sample can be determined by using a spectrophotometer? (3 pts)
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SPECTROPHOTOMETRIC ANALYSIS OF NUCLEIC ACIDS Mehmet Cetin Fall - 2008
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Objectives Learn how to do dilutions Use a spectrophotometer to calculate concentrations of nucleic acids Use absorbance data to figure out the purity of nucleic acids
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Spectroscopy Nucleic acids absorb light Best absorbance at 260nm
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Absorbance and Concentration Different molecules have different absorbance properties Absorbance reading of 1.0: dsDNA: 50 ug/ml ssDNA or RNA: 40 ug/ml Single stranded oligonucleotides: 33 ug/ml
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280nm: absorbance peak for proteins A260 / A280 ratio for purity 1.8 to 1.9 indicate high quality DNA. Low ratios indicate proteins 230nm: absorbance peak for phenol A260 / A230 ratio for purity. The lower the less pure. A325: contamination from cuvette. Should be 0
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This note was uploaded on 09/12/2009 for the course BISC 320L taught by Professor Baker,aparicio during the Fall '07 term at USC.

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Lab 1 SPECTROPHOTOMETRIC ANALYSIS OF NUCLEIC ACIDS - Office...

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