{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"volume":["21(4)"],"submitter":["Moazzam Jazi M"],"pubmed_abstract":["The quality and quantity of RNA are critical for successful downstream transcriptome-based studies such as microarrays and RNA sequencing (RNA-Seq). RNA isolation from woody plants, such as Pistacia vera, with very high amounts of polyphenols and polysaccharides is an enormous challenge. Here, we describe a highly efficient protocol that overcomes the limitations posed by poor quality and low yield of isolated RNA from pistachio and various recalcitrant woody plants. The key factors that resulted in a yield of 150 μg of high quality RNA per 200 mg of plant tissue include the elimination of phenol from the extraction buffer, raising the concentration of β-mercaptoethanol, long time incubation at 65 °C, and nucleic acid precipitation with optimized volume of NaCl and isopropyl alcohol. Also, the A260/A280 and A260/A230 of extracted RNA were about 1.9-2.1and 2.2-2.3, respectively, revealing the high purity. Since the isolated RNA passed highly stringent quality control standards for sensitive reactions, including RNA sequencing and real-time PCR, it can be considered as a reliable and cost-effective method for RNA extraction from woody plants."],"journal":["Physiology and molecular biology of plants : an international journal of functional plant biology"],"pagination":["597-603"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC4646865"],"repository":["biostudies-literature"],"pubmed_title":["Isolation of high quality RNA from pistachio (Pistacia vera L.) and other woody plants high in secondary metabolites."],"pmcid":["PMC4646865"],"pubmed_authors":["Seyedi SM","Moazzam Jazi M","Rajaei S"],"additional_accession":[]},"is_claimable":false,"name":"Isolation of high quality RNA from pistachio (Pistacia vera L.) and other woody plants high in secondary metabolites.","description":"The quality and quantity of RNA are critical for successful downstream transcriptome-based studies such as microarrays and RNA sequencing (RNA-Seq). RNA isolation from woody plants, such as Pistacia vera, with very high amounts of polyphenols and polysaccharides is an enormous challenge. Here, we describe a highly efficient protocol that overcomes the limitations posed by poor quality and low yield of isolated RNA from pistachio and various recalcitrant woody plants. The key factors that resulted in a yield of 150 μg of high quality RNA per 200 mg of plant tissue include the elimination of phenol from the extraction buffer, raising the concentration of β-mercaptoethanol, long time incubation at 65 °C, and nucleic acid precipitation with optimized volume of NaCl and isopropyl alcohol. Also, the A260/A280 and A260/A230 of extracted RNA were about 1.9-2.1and 2.2-2.3, respectively, revealing the high purity. Since the isolated RNA passed highly stringent quality control standards for sensitive reactions, including RNA sequencing and real-time PCR, it can be considered as a reliable and cost-effective method for RNA extraction from woody plants.","dates":{"release":"2015-01-01T00:00:00Z","publication":"2015 Oct","modification":"2024-10-18T00:05:11.56Z","creation":"2019-03-27T02:02:02Z"},"accession":"S-EPMC4646865","cross_references":{"pubmed":["26600686"],"doi":["10.1007/s12298-015-0319-x"]}}