| 余乃通1，周启林1，罗志文2，胡福初2，张治礼2，刘志昕1.仙人掌总RNA 提取方法的改进与分析[J].广东农业科学,2017,44(3):75-79 |
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仙人掌总RNA 提取方法的改进与分析 |
| Improvement and analysis of cactus totalRNA extraction method |
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| DOI：10.16768/j.issn.1004-874X.2017.03.011 |
| 中文关键词: 仙人掌 RNA 提取 RT-PCR 灵敏度检测 |
| 英文关键词: cactus RNA extraction RT-PCR sensitivity detection |
| 基金项目:海南省热带果树生物学重点实验室开放课题（KFZX2017002）；海南省自然科学基金（20153130） |
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| 中文摘要: |
| 利用亚硫酸钠分别与TRIzol 试剂、RNA plant plus Reagent 试剂、BioTeke 多糖多酚植物总RNA
提取试剂、艾德莱EASY spin Plus 植物RNA 快速提取试剂和OMEGA Plant RNA 试剂结合的方法，均可获
得高质量的仙人掌茎总RNA。反转录成cDNA，RT-PCR 结果显示，5 种方法提取的总RNA 反转录后均能
特异性扩增到matK 基因。使用TRIzol 试剂盒、RNA plant plus Reagent 试剂盒和艾德莱EASY spin Plus 植
物RNA 快速提取试剂盒提取RNA 质量相对较高，在cDNA 以1∶100 稀释时能检测到matK 基因；而使用
BioTeke 多糖多酚植物总RNA 提取试剂盒或OMEGA Plant RNA 试剂盒时，灵敏度稍低，在cDNA 以1∶10
稀释时，能检测到目的基因。利用亚硫酸钠结合不同植物总RNA 提取试剂盒进行比较研究，为后续制备仙
人掌茎高质量cDNA 文库及其分子生物学研究奠定基础。 |
| 英文摘要: |
| High-quality total RNAs of cactus stem were obtained by using Na2SO3 with each of TRIzol reagent，
RNA plant plus reagent，BioTeke total RNA extraction reagent，EASY spin Plus plant RNA extraction reagent
（Aidlab） or OMEGA plant RNA reagent. The cDNA RT-PCR and specific detection results showed that matK gene
fragments were amplified by five total RNA extraction methods. The cDNAs that reverse transcribed from the TRIzol
reagent，RNA Plant Plus Reagent，and EASY spin Plus Plant RNA Extraction reagent（ Aidlab） were relatively
high，as the sensitivity PCR of cDNA highest dilution to 1∶100. While the cDNAs that reverse transcribed from
BioTeke total RNA extraction reagent and OMEGA plant RNA reagent were relatively low，as the sensitivity RTPCR
indicated the cDNA highest dilution to 1∶10. In this study，Na2SO3 combined with different plant total RNA
extraction kits to prepare high quality of RNA is the key of cactus stem cDNA library preparation and molecular
biology. |
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