bisc320-lab1 - Shum 1 Alice Shum Section 13311 Lab Partners...

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Alice Shum Section # 13311 Lab Partners: Amy Cao, Date: 9/08/2010 Lab 1: Spectrophotometric Analysis of Nucleic Acids and DNA Agarose Gel Electrophoresis Introduction: The main objective of this experiment is to perform and calculate dilutions. Through this experiment, we learn to use a spectrophotometer to analyze the purity of single-stranded RNA and double-stranded DNA. Another objective is to determine the concentration of the nucleic acids. Then, we run the agarose gel electrophoresis to study diluted samples of RNA and DNA. The theory used in this experiment was the principles of spectrophotometry in that with a light source would create diffraction gratings. The light beam will penetrate through the cuvet with the sample solution to the photoelectric tube where the galvanometer will read the transmittance and convert it to an absorbance value. A new technique introduced was using the agarose for gel electrophoresis. This gel electrophoresis works through the movement of electrically charged particles of the sample in an electric field. Operating by the separation of macromolecules, the electrophoresis technique depends on the size with shape and charge density constant. The fragments must be stain with fluorescent dye, like ethidium bromide, to visualize the DNA fragments. Also, a new instrumental method used is the spectrophotometer called BioMate TM 3 UV spec-visible spectrophotometer. This instrument will allow us to set the wavelength desired to measure a blank and sample in order to get the absorbance. By performing this experiment, we expect to accomplish to learn that A 260 cannot differentiate between RNA and DNA. We also discover that the purity of the nucleic acid can be determined by the A 260 :A 280 ratio and wavelength A 230 to A 325 readings. Procedural Outline: Part 1: Dilutions and Spectrophotometry 2 samples will be given to each group in which one is double-strand DNA and the other is single-strand RNA. Then, take all samples and make a set of dilutions referring to Table 1
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This note was uploaded on 09/21/2010 for the course ALI 231 taught by Professor Staff during the Fall '08 term at USC.

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bisc320-lab1 - Shum 1 Alice Shum Section 13311 Lab Partners...

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