title: Automated serial extraction of DNA and RNA from biobanked tissue specimens.
abstract: BACKGROUND
  With increasing biobanking of biological samples, methods for large scale extraction of nucleic acids are in demand. The lack of such techniques designed for extraction from tissues results in a bottleneck in downstream genetic analyses, particularly in the field of cancer research. We have developed an automated procedure for tissue homogenization and extraction of DNA and RNA into separate fractions from the same frozen tissue specimen. A purpose developed magnetic bead based technology to serially extract both DNA and RNA from tissues was automated on a Tecan Freedom Evo robotic workstation.
  
  
  RESULTS
  864 fresh-frozen human normal and tumor tissue samples from breast and colon were serially extracted in batches of 96 samples. Yields and quality of DNA and RNA were determined. The DNA was evaluated in several downstream analyses, and the stability of RNA was determined after 9 months of storage. The extracted DNA performed consistently well in processes including PCR-based STR analysis, HaloPlex selection and deep sequencing on an Illumina platform, and gene copy number analysis using microarrays. The RNA has performed well in RT-PCR analyses and maintains integrity upon storage.
  
  
  CONCLUSIONS
  The technology described here enables the processing of many tissue samples simultaneously with a high quality product and a time and cost reduction for the user. This reduces the sample preparation bottleneck in cancer research. The open automation format also enables integration with upstream and downstream devices for automated sample quantitation or storage.
authors: Science for Life Laboratory, Department of Immunology, Genetics and Pathology, Uppsala University, SE-751 85, Uppsala, Sweden.
authors: Lucy
authors: L
authors: Mathot
authors: Monica
authors: M
authors: Wallin
authors: Tobias
authors: T
authors: Sjöblom
mesh_terms: D018070:Biological Specimen Banks; D004247:DNA / Q000302:isolation & purification*; D004273:DNA, Neoplasm / Q000302:isolation & purification; D018628:Gene Dosage; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D006801:Humans; D046228:Microarray Analysis / Q000379:methods; D009369:Neoplasms / Q000235:genetics; D011786:Quality Control; D012313:RNA / Q000302:isolation & purification*; D012334:RNA, Neoplasm / Q000302:isolation & purification; D015203:Reproducibility of Results; D012371:Robotics
chemical_list: D004273:DNA, Neoplasm; D012334:RNA, Neoplasm; D012313:RNA; D004247:DNA
references: Eur J Cancer. 2006 Nov;42(16):2684-91
references: 17027255
references: Chem Commun (Camb). 2011 Jan 7;47(1):547-9
references: 21103565
references: Biotechniques. 1995 Jan;18(1):62-3
references: 7702855
references: Science. 2006 Oct 13;314(5797):268-74
references: 16959974
references: Lab Invest. 2006 Feb;86(2):202-11
references: 16402036
references: PLoS One. 2012;7(12):e52750
references: 23300761
references: Proc Natl Acad Sci U S A. 2007 May 29;104(22):9387-92
references: 17517648
references: Clin Cancer Res. 2008 Dec 15;14(24):8198-204
references: 19088036
references: BMC Mol Biol. 2006 Jan 31;7:3
references: 16448564
references: Clin Chem. 2011 Nov;57(11):1545-55
references: 21914789
references: Methods Mol Biol. 2011;675:405-13
references: 20949406
references: Nature. 2008 Nov 6;456(7218):53-9
references: 18987734
references: J Korean Soc Coloproctol. 2010 Oct;26(5):316-23
references: 21152133