METHODS article
Front. Genet.
Sec. Genomic Assay Technology
Volume 16 - 2025 | doi: 10.3389/fgene.2025.1608504
A partially automated method for DNA extraction from marmoset hair follicles to avoid blood chimerism
Provisionally accepted
Donald Conrad1*
Alexandra M Stendahl1
Qiangge Zhang2Ana C Lima1Curtis Mello3,4James Nemesh3,4Sam Peterson1Jenna Castro1
Karina Ray1Xian Gao2Yuanyuan Hou2
Chenjie Shen2Katinka A Vigh-Conrad1Fenna Krienen5
Guoping Feng2Steven A Mccarroll3,4Ricardo C H Del Rosario3,4- 1Oregon National Primate Research Center, Oregon Health & Science University, Beaverton, United States
- 2McGovern Institute for Brain Research, School of Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States
- 3Stanley Center for Psychiatric Research, Broad Institute, Cambridge, Maryland, United States
- 4Department of Genetics, Harvard Medical School, Boston, Massachusetts, United States
- 5Princeton Neuroscience Institute, Princeton University, Princeton, New Jersey, United States
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Marmosets are valuable non-human primate models, however their unique reproduction results in high levels of blood chimerism, making blood unreliable for DNA sequencing. Hair follicles have lower levels of chimerism however DNA extraction from hair follicles is challenging due to the limited tissue. We developed a non-invasive, partially automated protocol for hair follicle collection and DNA extraction scalable to hundreds of samples. This method uses a proteinase K cell lysis solution in conjunction with Promega's Maxwell RSC's paramagnetic silica-based particles to purify DNA. We applied this protocol to samples collected from over 300 animals and from two different species. We were able to generate high quality libraries for whole genome sequencing (WGS) from approximately 150 hair follicles. Libraries built from >0.15µg DNA had an average duplication rate of 0.19, analogous to libraries built from blood. Sequenced DNA had average chimerism rates of 2.3%. DNA extraction from hair follicles offers a reliable method for whole genome sequencing with minimal chimerism. The partial automation improves efficiency by reducing lab time and extraction variability. The protocol is applicable to a range of projects requiring low-input DNA sources or automated, high-throughput sample processing.
Keywords: hair follicles, DNA extraction, Maxwell RSC, Chimerism, marmoset
Received: 09 Apr 2025; Accepted: 02 Sep 2025.
Copyright: © 2025 Conrad, Stendahl, Zhang, Lima, Mello, Nemesh, Peterson, Castro, Ray, Gao, Hou, Shen, Vigh-Conrad, Krienen, Feng, Mccarroll and Del Rosario. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Donald Conrad, Oregon National Primate Research Center, Oregon Health & Science University, Beaverton, United States
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