Amendolara, Alfred

Bills, Kyle B

Dougherty, John, Jr.

Farnsworth, Cole

Guzman, Steve

Henley, Molly

Hill, Nathaniel E

Kriak, John A

Minor, Laura

Reynolds, Jonathon

Sant, David W

Shah, Mujtaba

English

eCommons at Roseman University

20/G 10223Qiagen Blood & Cell Culture DNA Mini Kit (25) 13323SkullThyroidSalivary glandTracheaThymusRibsBile ductPancreasEsophagusAortic archLungLiverStomachKidneySpleenIleumIntestineBladderThyroidSalivary glandTracheaThymusRibsBile ductPancreasAortic archHeartLungLiverStomachKidneySpleenIleumIntestineBladderOvaryCervixBrainSpinalColumnBladderComparison of DNA Extraction Methods for Soft Tissue SamplesSteve Cochancela Guzman*, Nathaniel Hill*, Mujtaba H. Shah, Jonathon Reynolds, Cole Farnsworth, Molly Henley, Alfred Amendolara,Laura Minor, John Jr. Dougherty, John A. Kriak, Kyle B. Bills, David W. Sant1Department of Biomedical Sciences, Noorda College of Osteopathic Medicine, Provo, UT* Authors Contributed EquallyTesting of somatic mutations and epigenetic DNA modifications requires extracting DNA from tissues. However, several substances present in tissues are known to inhibit PCR, and commercial kits are traditionally not designed to work with all tissues. We will attempt to determine which commercial DNA extraction kits will yield high-molecular weight DNA that is free of PCR inhibitors.BackgroundFigure 3. HbA1c scoresMethodsPCR Inhibitors:Myoglobin FatsHemoglobin CalciumBilirubin MagnesiumAntibodies Bile SaltsHeparin UreaCollagen MelaninLactoferrin HormonesPolysaccharidesOrgans Tested:Heart SkinLiver LungsSpleen KidneysAdrenal GlandsTesticlesBone MarrowSkeletal MuscleTissues from 48 Sprague-Dawley rats were collected. Post-mortem rats from a morphine study were used to lessen animal burden. Homogenized organs were aliquoted at 10 mg. qPCR, Qubit fluorometer, and gel electrophoresis will be used to evaluate extracted DNA for inhibitors, amount, and fragment length.Picture 2: Anatomy of a rat demonstrating vital organs adapted from Science Photo Library Picture 1: Tissue homogenizer similar tothe one used for homogenization of organs00.511.522.533.544.5Heart Lungs Pancreas Spleen Kidneys AdrenalGlandsTesticles RetinaOrgan WeightsMale FemaleFigure 1: Weight of organs gathered from rats (grams)Figure12. adapted from Young et al. 2015 Annu Rev Nutr. Various cytosine modifications change with tissues and in disease states, necessitating extraction of DNA directly from tissues. Table 1: Kits ordered for extraction of DNA from various solid tissuesPicture 2: Instrumentation used to test quality of DNAEsophagusIleumHeart

Comparison of DNA Extraction Methods for Soft Tissue Samples

https://ecommons.roseman.edu/cgi/viewcontent.cgi?article=1297&context=researchsymposium

Comparison of DNA Extraction Methods for Soft Tissue Samples

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