Ancient DNA: Methods and Protocols by Tara L. Fulton (auth.), Beth Shapiro, Michael Hofreiter

By Tara L. Fulton (auth.), Beth Shapiro, Michael Hofreiter (eds.)

Research into historic DNA started greater than 25 years in the past with the e-book of brief mitochondrial DNA series fragments from the quagga, an extinct relative of the zebra. historic DNA examine quite won momentum following the discovery of PCR, which allowed thousands of copies to be made from the few last DNA molecules preserved in fossils and museum specimens. In Ancient DNA: equipment and Protocols professional researchers within the box describe some of the protocols which are now customary to check historical DNA. those comprise directions for establishing an historic DNA laboratory, extraction protocols for quite a lot of diverse substrates, info of laboratory strategies together with PCR and NGS library education, and proposals for acceptable analytical methods to make experience of the sequences received. Written within the hugely winning Methods in Molecular Biology™ sequence layout, chapters comprise introductions to their respective themes, lists of the required fabrics and reagents, step by step, quite simply reproducible laboratory protocols, and key tips about troubleshooting and warding off identified pitfalls.

Authoritative and sensible, Ancient DNA: equipment and Protocols seeks to help scientists within the additional research of historical DNA and the methodological ways in historical research.

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Store in the dark at +4°C. 9. Prior to use, vortex to resuspend any pelleted material. 2. Paleofeces DNA Extraction 1. Add approximately 1 g of fecal material to a small weighing boat (see Note 1). 2. Cut the fecal remains into small pieces using scalpel blades (see Note 2). 40 M. Kuch and H. Poinar 3. Add fecal material to a final volume of 14 mL of the GuSCN extraction-buffer in a 15-mL tube and incubate, rotating overnight at 37°C in the dark (see Notes 1, 3 and 4). 4. Centrifuge at maximum speed for 5 min and transfer supernatant to a clean 15-mL tube.

5)). 6. 05% Tween-20 (60 μL per sample). 7. Glacial Acetic acid (~15 μL per sample). 3. 1. Preparing the Silica Suspension 1. 8 g silica and place it into a 50-mL gammasterilized tube. 2. Add double-distilled (dd) H2O to the tube containing the silica to 40 mL, vortex for 2 min, then let sit for 24 h at room temperature in the dark. 3. Carefully remove 35 ml of supernatant (without distrurbing the pellet) and discard. 4. Add ddH2O to 40 mL, vortex for 2 min, let sit for 6 h at room temperature in the dark.

Proc Biol Sci 276:3395 4. Willerslev E, Gilbert MT, Binladen J et al (2009) Analysis of complete mitochondrial genomes from extinct and extant rhinoceroses reveals lack of phylogenetic resolution. BMC Evol Biol 9:95 5. King G, Gilbert M, Willerslev E et al (2009) Recovery of DNA from archaeological insect remains: first results, problems and potential. J Archaeol Sci 36:1179–1183 6. Yang DY, Eng B, Waye JS et al (1998) Technical note: improved DNA extraction from ancient bones using silica-based spin columns.

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