title: Storage by lyophilization - Resulting RNA quality is tissue dependent.
abstract: In today's highly collaborative scientific community there is a growing need to transport biological samples across the globe. Lyophilization is a cost-effective preservation method which avoids the use of hazardous chemicals, creating an appealing, yet essentially unexplored, prospect for the long-range transport of animal tissue samples. This study examined the integrity of RNA following its extraction from eel tissue (liver, spleen and ovary) that had been subjected to i) freezing only; ii) freezing and lyophilization, and iii) freezing, lyophilization and subsequent storage at ambient temperature for one week. Only small reductions in RNA integrity were identified in lyophilized, stored sample compared to that of flash-frozen or lyophilized sample not subjected to storage. Reductions in RNA integrity were most profound in ovary tissue, which has a notably higher lipid content (∼35% of dry weight) than liver (∼17%) or spleen (∼15%). However, lowered RNA integrity numbers did not affect qPCR-estimated relative or absolute transcript copy numbers of two arbitrary target genes. These findings confirm that this method is a viable option for shipping animal tissue samples world-wide and could open up numerous benefits for the biological sciences as a whole.
authors: Department of Zoology, University of Otago, PO Box 56, Dunedin 9054, New Zealand. Electronic address: erin.damsteegt@otago.ac.nz.
authors: Erin L
authors: EL
authors: Damsteegt
authors: Department of Zoology, University of Otago, PO Box 56, Dunedin 9054, New Zealand.
authors: Nicky
authors: N
authors: McHugh
authors: Department of Zoology, University of Otago, PO Box 56, Dunedin 9054, New Zealand.
authors: P Mark
authors: PM
authors: Lokman
mesh_terms: D000818:Animals; D004524:Eels; D005260:Female; D005612:Freeze Drying; D008099:Liver / Q000737:chemistry*; D009928:Organ Specificity; D010053:Ovary / Q000737:chemistry*; D012313:RNA / Q000737:chemistry* / Q000302:isolation & purification*; D013154:Spleen / Q000737:chemistry*
chemical_list: D012313:RNA
keywords: Freeze-dried; Lyophilization; RIN number; RNA quality; RNA storage; qPCR