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

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

Research into historical DNA started greater than 25 years in the past with the book of brief mitochondrial DNA series fragments from the quagga, an extinct relative of the zebra. historical DNA study fairly won momentum following the discovery of PCR, which allowed hundreds of thousands of copies to be made up of the few final DNA molecules preserved in fossils and museum specimens. In Ancient DNA: tools and Protocols professional researchers within the box describe a few of the protocols which are now usual to review historical DNA. those contain directions for developing an historic DNA laboratory, extraction protocols for a variety of varied substrates, info of laboratory ideas together with PCR and NGS library guidance, and recommendations for acceptable analytical techniques to make experience of the sequences got. Written within the hugely winning Methods in Molecular Biology™ sequence layout, chapters comprise introductions to their respective subject matters, lists of the mandatory fabrics and reagents, step by step, effectively reproducible laboratory protocols, and key pointers on troubleshooting and fending off identified pitfalls.

Authoritative and functional, Ancient DNA: tools and Protocols seeks to help scientists within the additional research of old DNA and the methodological methods in old research.

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5. 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.

Rohland N, Hofreiter M (2007) Comparison and optimization of ancient DNA extraction. Biotechniques 42:343–352 9. Poinar HN (2002) The genetic secrets some fossils hold. Acc Chem Res 35:676–684 10. Vasan S, Zhang X, Kapurnitou A, Bernhagen J, Teichberg S, Basgen J, Wagle D, Shih D, Terlecky I, Bucala R, Cerami A, Egan J, Uhlrich P (1996) An agent cleaving glucose-derived protein crosslinks in vitro and in vivo. Nature 382:275–278 11. Adler CJ, Haak W, Donlon D, Cooper A (2010) Survival and recovery of DNA from ancient teeth and bones.

Always handle both liquids and their containers with extreme care, using appropriate face, hand, and body protection. Do not handle using latex gloves, as these are permeable to phenol and chloroform; use only nitryl gloves. The fumes of both chemicals are dangerous; therefore, these steps should always be performed in a vented fume hood. Disposal of both requires conformation to specific regulations, thus relevant local disposal regulations should be consulted. 9. 5-mL tubes or smaller are used).

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