title: A lysis, storage, and transportation buffer for long-term, room-temperature preservation of human clinical lymphoid tissue samples yielding high molecular weight genomic DNA suitable for molecular diagnosis.
abstract: Molecular diagnosis of malignant lymphoma depends on the ability to extract high molecular weight genomic DNA. However, collection, storage, and transportation of frozen tissue is time consuming and expensive. We used a simple, low-cost lysis, storage, and transportation buffer (LST) to maintain clinical tissue samples at room temperature for up to 4 weeks before molecular analysis. Immersion of lymphoid tissue in LST at room temperature for 2 to 4 weeks was compared with snap-freezing in liquid nitrogen followed by storage at -75 degrees C. Southern blot analysis using an immunoglobulin heavy chain JH probe yielded identical results in 5 clonal and 6 nonclonal samples. The DNA recovered from the LST of a 12th sample was too degraded to be analyzed; however, the tissue had large zones of geographic necrosis. We also demonstrated that DNA extracted from tissue stored in LST is suitable for amplification by the polymerase chain reaction. Results from 4 of the snap-frozen and LST samples analyzed for rearrangements at the immunoglobulin heavy chain VDJ locus were identical. LST can be used in a clinical laboratory for storing tissue samples at room temperature up to 4 weeks before molecular analysis.
authors.affiliation: Department of Pathology, Albert Einstein College of Medicine-Montefiore Medical Center, Bronx, NY 10467, USA.
authors.forename: C L
authors.initials: CL
authors.lastname: Schultz
authors.forename: Y
authors.initials: Y
authors.lastname: Akker
authors.forename: J
authors.initials: J
authors.lastname: Du
authors.forename: H
authors.initials: H
authors.lastname: Ratech
mesh_terms: D015139:Blotting, Southern; D003582:Cytogenetics / Q000379:methods*; D004247:DNA / Q000302:isolation & purification*; D004586:Electrophoresis; D015321:Gene Rearrangement; D005803:Genes, Immunoglobulin; D006801:Humans; D008221:Lymphoid Tissue / Q000166:cytology*; D008970:Molecular Weight; D011092:Polyethylene Glycols; D016133:Polymerase Chain Reaction; D011136:Polysorbates; D014021:Tissue Preservation / Q000379:methods*; D014325:Tromethamine
chemical_list: C119344:LST buffer; D011136:Polysorbates; D014325:Tromethamine; D011092:Polyethylene Glycols; D004247:DNA