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首页> 外文期刊>Preparative biochemistry & biotechnology: An international journal for rapid communication >Efficient Extraction of High-Quality Total RNA from Various Hop Tissues (Humulus lupulus L.)
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Efficient Extraction of High-Quality Total RNA from Various Hop Tissues (Humulus lupulus L.)

机译：从各种蛇麻草组织中高效提取高质量的总RNA

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摘要

The isolation of high-quality RNA is a prerequisite for gene expression studies.RNA quality is of special relevance if PCR-based strategies such as cDNA-AFLP or RT-PCR are followed.Our molecular investigations of hop (Humulus lupulus L.) focus on genes that determine the biosynthesis of prenylflavonoids with interesting biological activities which accumulate in the lupulin glands.However,optimized protocols for RNA extraction from hop cones are not available.In this study,the RNeasy midi kit protocol was modified to isolate high amounts of total RNA from fresh and freeze-dried hop tissues,specifically leaves,female inflorescences,and lupulin-enriched hop cone fractions.The main difficulties in obtaining high RNA yields were related to specific features of hop,including the abundance of secondary metabolites and their accumulation in the sticky lupulin glands.The introduction of a number of modifications into the RNeasy midi kit protocol resulted in RNA of high quality,as assessed by spectrophotometry and electrophoresis in agarose gels.The protocol developed is currently being used to produce cDNA-AFLP fingerprints of hop cones and leaves for genetic screening.Furthermore,the method could provide RNA for further molecular studies,including Northern blot hybridizations and reverse transcription-polymerase chain reactions.
机译：高质量R​​NA的分离是进行基因表达研究的先决条件。如果遵循基于PCR的策略（例如cDNA-AFLP或RT-PCR），则RNA的质量特别重要。我们对蛇麻草（Humulus lupulus L.）进行分子研究能够确定具有重要生物学活性的异戊二烯类黄酮生物合成的基因，这些异戊二烯类黄酮类化合物在卢普林腺体中积累。但是，尚无从啤酒花球果中提取RNA的优化方案。在本研究中，对RNeasy midi试剂盒方案进行了修改，以分离出大量总RNA。来自新鲜和冻干啤酒花组织的RNA，特别是叶片，女性花序和富含卢普林的啤酒花锥组分。获得高RNA产量的主要困难与啤酒花的特定特征有关，包括次生代谢产物的丰度及其在啤酒花中的积累黏附的叶绿素腺体。如通过sp评估，在RNeasy midi试剂盒规程中引入了许多修饰产生了高质量的RNA琼脂糖凝胶中的分光光度法和电泳。目前开发的方案可用于生产啤酒花球果和叶片的cDNA-AFLP指纹以进行遗传筛选。此外，该方法可为RNA的进一步研究提供分子，包括RNA印迹杂交和逆转录聚合酶连锁反应。

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