BMI_lab9_Quantitation

BMI_lab9_Quantitation - Quantitation of DNA by UV...

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Quantitation of DNA by UV spectrophotometry and plasmid restriction digest Laboratory Exercise 9
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Objective To determine the quantity and quality of pUC19 DNA isolated from transformed cell culture To perform a restriction digest of pUC19 DNA To perform a simulated restriction digest of λ DNA
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Quantitation of DNA The quantitation of DNA will be done using a spectrophotometer, similar to what was done in laboratory 2. Recall: Absorbance is proportional to the concentration. Also known as OD
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Quantitation of DNA The wavelength of light measured depends on the target molecule DNA and RNA will absorb light maximally at 260nm Proteins will absorb maximally at 280nm
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Quantitation of DNA An OD 260 of 1 corresponds to a concentration of 50 ug/ml for double stranded DNA, This is same as 50ng/ul of DNA
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Quantitation of DNA A ratio of OD 260nm/280 measures the purity of your sample For Pure DNA the ratio of OD 260/280 is between 1.8-2.0 Examples:
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This note was uploaded on 04/24/2008 for the course PHS 2301 taught by Professor Carvalho during the Spring '08 term at St. Johns Duplicate.

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BMI_lab9_Quantitation - Quantitation of DNA by UV...

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