EXTRACTING DNA FROM C ELEGANS By Brianna N

EXTRACTING DNA FROM C. ELEGANS By: Brianna N. Kaylor, Joseph H. Lackey And Ryan Kelly

C. elegans 1, 582 base pairs for C. elegans organic anion transporter Intestine 2111136 www. wormbook. org

Extracting the DNA � � Lyse C. elegans Bind DNA to Column Wash Elute DNA from Column Detect DNA Extract DNA C. elegans Spectroscopic quantification Agarose gel electrophoresis detection www. wormbook. org

Techniques Gel Electrophoresis Spectrophotometer Can analyze a 1 μL sample of DNA Use an electromagnetic field to pull the DNA from the wells to create the bands that we can see on the UV imager.

Results DNA extract absorbance 14 12 A 260/A 280 = 1. 59 10 Absorbance units 338 ng/n. L 8 6 4 2 0 200 -2 220 240 260 280 wavelength (nm) 300 320 340

Analyzing the gel Agarose gel electrophoresis Poor separation of DNA molecules > 50 kb UV UV ethidium bromide dye is used to stain DNA

Results A single band stuck in the wells is whole, uncut genomic DNA. Solutions: 1)Obtain greater purity of DNA 2) Use lower agarose concentrations 3) Use enzymatic digestion to cut fragments into smaller pieces Marker DNA A smear of DNA in the lane represents large pieces of partially degraded genomic DNA,

Acknowledgements � � Academy of Science program Dr. Cecile A. R. Smith Department of Chemistry Dr. Tashakkori