The Criteria of Authentication

We tried to follow the instructions in Criteria of Authenticity (Hofreiter 2001). Briefly, in-laboratory procedures were performed in the rooms only devoted to aDNA work; any modern samples were by no means imported to the rooms. PCR reaction was repeated in our lab to show reproducibility of amplification, during which negative control was always used for excluding possible contamination of modern DNA during lab works.

Reference: Hofreiter, M., Serre, D., Poinar, H.N., Kuch, M. & Pääbo, S. (2001): Ancient DNA. - Nat. Rev. Genet. 2, 353-359.

Examples of Criteria of Authentication observed in my lab

Extraction controls and PCR controls: Mock extractions and PCRs without template DNA are elementary controls that should be carried out.

Inverse correlation between amplicon length and amplification efficiency: Generally, amplification of only short DNA pieces is possible.

Quantitation of numbers of template molecules: If the number of DNA molecules that initiate the PCR is less than ~1,000, at least three independent amplifications need to be analysed, and the products need to be cloned and several clones should be sequenced.

Amplification from a second extract: The reproduction of the results from a second, independent extract should show that the result is reliable.


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