title: Fasciola hepatica - where is 28S ribosomal RNA?
abstract: Advanced molecular biology techniques are currently used to develop new effective strategies against fasciolosis. Assessment of the quality of extracted total RNA is an important step prior to commencing many molecular biology methods such as transcriptomics. However, RNA quality assessment is complicated for some organisms, including Fasciola hepatica, by the absence of a 28S rRNA peak/band, when assessed with modern protocols. In this study, electrophoretic profiles of F. hepatica ribosomal RNAs were evaluated using microfluidics capillary based and conventional non-denaturing gel electrophoresis methods. An important modification to recommended protocols, the exclusion of heat-denaturation step, in the microfluidics capillary based electrophoresis is critical to visualise the expected 28S rRNA and obtain an RNA integrity number (RIN). The intensity of the 28S rRNA band is reduced by the effect of non-denaturing gel electrophoresis.
authors: TÜBİTAK Marmara Research Center, Genetic Engineering and Biotechnology Institute, P.O. Box 21, 41470 Gebze, Kocaeli, Turkey. Electronic address: orcun.hacariz@tubitak.gov.tr.
authors: Orçun
authors: O
authors: Haçarız
authors: Gearóid
authors: G
authors: Sayers
mesh_terms: D000818:Animals; D002417:Cattle; D004587:Electrophoresis, Agar Gel; D019075:Electrophoresis, Capillary / Q000379:methods / Q000592:standards; D005210:Fasciola hepatica / Q000235:genetics*; D006358:Hot Temperature; D044085:Microfluidics; D018510:RNA, Helminth / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D012335:RNA, Ribosomal / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D012337:RNA, Ribosomal, 18S / Q000737:chemistry / Q000302:isolation & purification; D012339:RNA, Ribosomal, 28S / Q000737:chemistry / Q000302:isolation & purification*; D012340:RNA, Ribosomal, 5.8S / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D018510:RNA, Helminth; D012335:RNA, Ribosomal; D012337:RNA, Ribosomal, 18S; D012339:RNA, Ribosomal, 28S; D012340:RNA, Ribosomal, 5.8S
keywords: 28S rRNA; Fasciola hepatica; Microfluidics capillary based electrophoresis; RIN number; RNA quality