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Squeezing water from a stone: high-throughput sequencing from a 145-year old holotype resolves (barely) a cryptic species problem in flying lizards. | LitMetric

AI Article Synopsis

  • Researchers successfully extracted and sequenced genetic data from a 145-year-old holotype of the flying lizard to solve a taxonomic issue regarding its species status among similar species.
  • Despite initial concerns about DNA degradation, a specialized extraction technique allowed them to obtain usable mitochondrial DNA for accurate analysis.
  • Their findings challenge the belief that formalin-fixed samples are the most difficult for sequencing, suggesting that ethanol-fixed specimens might face more significant degradation over time.

Article Abstract

We used Massively Parallel High-Throughput Sequencing to obtain genetic data from a 145-year old holotype specimen of the flying lizard, . Obtaining genetic data from this holotype was necessary to resolve an otherwise intractable taxonomic problem involving the status of this species relative to closely related sympatric species that cannot otherwise be distinguished from one another on the basis of museum specimens. Initial analyses suggested that the DNA present in the holotype sample was so degraded as to be unusable for sequencing. However, we used a specialized extraction procedure developed for highly degraded ancient DNA samples and MiSeq shotgun sequencing to obtain just enough low-coverage mitochondrial DNA (721 base pairs) to conclusively resolve the species status of the holotype as well as a second known specimen of this species. The holotype was prepared before the advent of formalin-fixation and therefore was most likely originally fixed with ethanol and never exposed to formalin. Whereas conventional wisdom suggests that formalin-fixed samples should be the most challenging for DNA sequencing, we propose that evaporation during long-term alcohol storage and consequent water-exposure may subject older ethanol-fixed museum specimens to hydrolytic damage. If so, this may pose an even greater challenge for sequencing efforts involving historical samples.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5865465PMC
http://dx.doi.org/10.7717/peerj.4470DOI Listing

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