6533b824fe1ef96bd1280c64
RESEARCH PRODUCT
How to preserve and handle fish liver samples to conserve RNA integrity
Eeva-riikka VehniäinenMaiju RuusunenAimo OikariPekka J. VuorinenMarja KeinänenJussi V. K. Kukkonensubject
MaleTroutBleached kraft pulp mill effluentHealth Toxicology and Mutagenesis010501 environmental sciencesSample storage01 natural sciencessäilytysSpecimen HandlingTranscriptomeBrown troutsample storageFish liverFreezingAnimalsEnvironmental Chemistryteollisuusjätevesi14. Life underwaterSalmoGene0105 earth and related environmental scienceskalatCryopreservationQuantitative reverse transcription PCRbiologyReverse Transcriptase Polymerase Chain ReactionsytokromitmaksaRNAnäytteetGeneral MedicineRna degradationbiology.organism_classificationPollutionbleached kraft pulp mill effluentPoikilothermLiverBiochemistrycytochrome p450quantitative reverse transcription PCRRNACytochrome p450TranscriptomeResearch ArticleRNA integritydescription
As transcriptomic studies are becoming more and more common, it is important to ensure that the RNA used in the analyses is of good quality. The RNA integrity may be compromised by storage temperature or freeze-thaw cycles, but these have not been well studied in poikilothermic fishes. This work studied the effects of tissue storage time and temperature, and freeze-thaw cycles of tissue and extracted RNA on RNA integrity in brown trout (Salmo trutta L.) liver. The storage time and temperature had an effect on RNA integrity, but RNA suitable for quantitative reverse transcription PCR (RT-qPCR) (RIN > 7) was still obtained from samples preserved at − 20 °C for 6 months. Freeze-thaw cycles of tissue or RNA did not compromise the integrity of RNA. RNA degradation had an effect on RT-qPCR results, and the effect depended on gene. The RT-qPCR analysis of historical samples from a bleached kraft pulp mill effluent exposure in 1984 revealed no significant cyp1a induction. Recommendations are given for the preservation and handling procedures of samples designated for transcriptomic analyses. peerReviewed
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2019-04-22 |