Identification and selection of optimal reference genes for qPCR-based gene expression analysis in Fucus distichus under various abiotic stresses
نویسندگان
چکیده
Quantitative gene expression analysis is an important tool in the scientist’s belt. The identification of evenly expressed reference genes necessary for accurate quantitative analysis, whether by traditional RT-PCR (reverse-transcription polymerase chain reaction) or qRT-PCR (quantitative real-time PCR; qPCR). In Stramenopiles (the major line eukaryotes that includes brown algae) there a noted lack known such studies, largely due to absence available molecular tools. Here we present set nine ( Elongation Factor 1 alpha (EF1A) , 2 (EF2A) beta (EF1B) 14-3-3 Protein Ubiquitin Conjugating Enzyme (UBCE2) Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH) Actin Related Complex (ARP2/3) Ribosomal (40s; S23) and ) alga Fucus distichus . These were tested on adult sporophytes across six abiotic stress conditions (desiccation, light temperature modification, hormone addition, pollutant exposure, nutrient wounding). Suitability these as was quantitatively evaluated using standard methods majority favorably. However, show normalization should be chosen condition-by-condition basis. We provide recommendation at least two used per experiment, list recommended pairs here, procedure identifying suitable experimenter’s unique design. With recent expansion interest algal biology accompanied tools development, variety experimental here makes this study valuable resource future work basic understanding responses lineage.
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ژورنال
عنوان ژورنال: PLOS ONE
سال: 2021
ISSN: ['1932-6203']
DOI: https://doi.org/10.1371/journal.pone.0233249