Improving Species Identification of Ancient Mammals Based on Next-Generation Sequencing Data

The taxonomical identification merely based on morphology is often difficult for ancient remains. Therefore, universal or specific PCR amplification followed by sequencing and BLAST (basic local alignment search tool) search has become the most frequently used genetic-based method for the species identification of biological samples, including ancient remains. However, it is challenging for these methods to process extremely ancient samples with severe DNA fragmentation and contamination. Here, we applied whole-genome sequencing data from 12 ancient samples with ages ranging from 2.7 to 700 kya to compare different mapping algorithms, and tested different reference databases, mapping similarities and query coverage to explore the best method and mapping parameters that can improve the accuracy of ancient mammal species identification. The selected method and parameters were tested using 152 ancient samples, and 150 of the samples were successfully identified. We further screened the BLAST-based mapping results according to the deamination characteristics of ancient DNA to improve the ability of ancient species identification. Our findings demonstrate a marked improvement to the normal procedures used for ancient species identification, which was achieved through defining the mapping and filtering guidelines to identify true ancient DNA sequences. The guidelines summarized in this study could be valuable in archaeology, paleontology, evolution, and forensic science. For the convenience of the scientific community, we wrote a software script with Perl, called AncSid, which is made available on GitHub

The complete mitochondrial genome of Melanoides tuberculata (Müller, 1774) in Guangdong, China

The Melanoides tuberculata is an invasive species, which is natively distributed in Africa and Southeast Asia. This study describes the first mitochondrial genome of the M. tuberculata based on the whole genome sequencing data. The complete sequence length of the mitogenome is 15,821 bp, including 37 genes (2 rRNA genes, 22 tRNA genes and 13 protein-coding genes). Phylogenetic analysis using the 13 species of Cerithioidea species showed that the M. tuberculata is closely related to P. dartevellei, forming the sister group to C. sinensis and C. obtuse

A simple Preservation Method of Bird Feces for DNA Analysis: A Case Study on Chicken and Quail

Standardized methods for fecal sample collection and safe long-distance transportation for DNA extraction are yet to be identified. We compared four different preservation methods for bird fecal samples: storage in 75% and 100% ethanol, freezing at −20°C, and immersing in 100% ethanol for 3 weeks followed by drying the samples for more than 60 days and transporting them to another country. Our objectives were to quantify the DNA concentration and amplify a fragment of the gene from each sample successfully using the primers mcb398 and mcb869 through DNA barcoding. Our data showed that the method of sample preservation used affected the DNA concentration and amplification. The best results were achieved when the samples were preserved in 100% ethanol. The samples were then dried for storage prior to further processing. This method is inexpensive and safe for long-distance transportation and at airports

An efficient pipeline for ancient DNA mapping and recovery of endogenous ancient DNA from whole-genome sequencing data

Abstract Ancient DNA research has developed rapidly over the past few decades due to improvements in PCR and next‐generation sequencing (NGS) technologies, but challenges still exist. One major challenge in relation to ancient DNA research is to recover genuine endogenous ancient DNA sequences from raw sequencing data. This is often difficult due to degradation of ancient DNA and high levels of contamination, especially homologous contamination that has extremely similar genetic background with that of the real ancient DNA. In this study, we collected whole‐genome sequencing (WGS) data from 6 ancient samples to compare different mapping algorithms. To further explore more effective methods to separate endogenous DNA from homologous contaminations, we attempted to recover reads based on ancient DNA specific characteristics of deamination, depurination, and DNA fragmentation with different parameters. We propose a quick and improved pipeline for separating endogenous ancient DNA while simultaneously decreasing homologous contaminations to very low proportions. Our goal in this research was to develop useful recommendations for ancient DNA mapping and for separation of endogenous DNA to facilitate future studies of ancient DNA