title: High-throughput, automated extraction of DNA and RNA from clinical samples using TruTip technology on common liquid handling robots.
abstract: TruTip is a simple nucleic acid extraction technology whereby a porous, monolithic binding matrix is inserted into a pipette tip. The geometry of the monolith can be adapted for specific pipette tips ranging in volume from 1.0 to 5.0 ml. The large porosity of the monolith enables viscous or complex samples to readily pass through it with minimal fluidic backpressure. Bi-directional flow maximizes residence time between the monolith and sample, and enables large sample volumes to be processed within a single TruTip. The fundamental steps, irrespective of sample volume or TruTip geometry, include cell lysis, nucleic acid binding to the inner pores of the TruTip monolith, washing away unbound sample components and lysis buffers, and eluting purified and concentrated nucleic acids into an appropriate buffer. The attributes and adaptability of TruTip are demonstrated in three automated clinical sample processing protocols using an Eppendorf epMotion 5070, Hamilton STAR and STARplus liquid handling robots, including RNA isolation from nasopharyngeal aspirate, genomic DNA isolation from whole blood, and fetal DNA extraction and enrichment from large volumes of maternal plasma (respectively).
authors.affiliation: Application Development, Akonni Biosystems, Inc. rholmberg@akonni.com
authors.forename: Rebecca C
authors.initials: RC
authors.lastname: Holmberg
authors.forename: Alissa
authors.initials: A
authors.lastname: Gindlesperger
authors.forename: Tinsley
authors.initials: T
authors.lastname: Stokes
authors.forename: Dane
authors.initials: D
authors.lastname: Brady
authors.forename: Nitu
authors.initials: N
authors.lastname: Thakore
authors.forename: Philip
authors.initials: P
authors.lastname: Belgrader
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Cooney
authors.forename: Darrell P
authors.initials: DP
authors.lastname: Chandler
mesh_terms: D004247:DNA / Q000097:blood / Q000302:isolation & purification*; D005260:Female; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D008297:Male; D017817:Nasal Lavage Fluid / Q000737:chemistry; D009305:Nasopharynx / Q000378:metabolism; D012313:RNA / Q000097:blood / Q000302:isolation & purification*; D012371:Robotics / Q000295:instrumentation / Q000379:methods
chemical_list: D012313:RNA; D004247:DNA