Korean pine （Pinus koraiensis Sieb. et Zucc.） is an ecologically and economically important tree species in East Asia. Molecular studies of seed development of this species are limited due to the lack of effective R...Korean pine （Pinus koraiensis Sieb. et Zucc.） is an ecologically and economically important tree species in East Asia. Molecular studies of seed development of this species are limited due to the lack of effective RNA extraction protocols. This study aimed to obtain an effec- tive method to extract high-quality RNA from Korean pine seeds. The TRIzol kit and CTAB methods were used to extract the total RNA from Korean pine seeds at different developmental stages. The bands of RNA extracted by CTAB were not clear, whereas the bands of RNA extracted by the TRIzol kit were brighter and clearer, indicating higher quality and integrity of the RNA products extracted by the TRIzol kit. The 28S rRNA band was approximately 1.5- to 2-fold brighter than the 18S rRNA band on the agarose gel electrophoresis. The absorbance value A 260/280 was 1.8-2.0, and the absorbance value A 260/230 was 〉1.9. The Bioanalyzer RNA integrity results showed that the RNA integrity number of the RNA extracted using the TR/zol kit was acceptable for high-throughputsequencing. Therefore, the total RNA extracted using the TRIzol kit method can be used for high-throughput sequencing and other molecular biology experiments.展开更多
副溶血弧菌(Vibrio parahaemolyticus)是一种革兰氏阴性嗜盐菌。提取高质量的RNA是研究副溶血弧毒力基因表达与其致病性和环境适应性的基础。本研究分别用SDS法、Trizol法,PureLinkTM RNA Mini Kit和Uniq-10柱式Trizol总RNA抽提试剂盒...副溶血弧菌(Vibrio parahaemolyticus)是一种革兰氏阴性嗜盐菌。提取高质量的RNA是研究副溶血弧毒力基因表达与其致病性和环境适应性的基础。本研究分别用SDS法、Trizol法,PureLinkTM RNA Mini Kit和Uniq-10柱式Trizol总RNA抽提试剂盒提取六株副溶血弧菌的总RNA,用普通琼脂糖凝胶电泳和核酸测定仪检测RNA的质量,并用RT-PCR法对四种提取方法进行比较。研究结果表明,Trizol法和Pure-LinkTM RNA Mini Kit简单快捷,提取的总RNA完整性好,纯度高,可用于后续实验的分子生物学研究;而Trizol法更适用于普通实验室细菌RNA的提取。展开更多
为建立草莓果实总RNA的提取方法,针对草莓成熟果实中富含多糖、多酚和色素等次级代谢物质,总RNA提取难度大的特点,比较改良的EASYspin植物RNA提取试剂盒法和改良的CTAB法提取草莓果实中总RNA的质量。通过琼脂糖凝胶电泳和核酸蛋白测定仪...为建立草莓果实总RNA的提取方法,针对草莓成熟果实中富含多糖、多酚和色素等次级代谢物质,总RNA提取难度大的特点,比较改良的EASYspin植物RNA提取试剂盒法和改良的CTAB法提取草莓果实中总RNA的质量。通过琼脂糖凝胶电泳和核酸蛋白测定仪(Nano Drop 2000)检测2种方法提取总RNA的浓度、纯度及完整性等。改良的EASYspin植物RNA提取试剂盒法和改良的CTAB法都能够完成草莓果实总RNA的提取,电泳检测在28S和18S处呈2条清晰的条带,OD260/OD280值和OD260/OD230值均在2.0左右;改良的EASYspin植物RNA提取试剂盒法成本较高,且提取总RNA质量劣于改良的CTAB法,但是EASYspin植物RNA提取试剂盒法操作简便,安全可靠,节省时间。通过RT-PCR验证,2种方法所获得的草莓果实总RNA质量较好,可达到分子生物学试验对RNA质量的要求。展开更多
基金supported by the Special Fund for Forest Scientific Research in the Public Welfare(No.201404320)the Youth Scientific Research Fund from Qiqihar University(2012 kM22)
文摘Korean pine （Pinus koraiensis Sieb. et Zucc.） is an ecologically and economically important tree species in East Asia. Molecular studies of seed development of this species are limited due to the lack of effective RNA extraction protocols. This study aimed to obtain an effec- tive method to extract high-quality RNA from Korean pine seeds. The TRIzol kit and CTAB methods were used to extract the total RNA from Korean pine seeds at different developmental stages. The bands of RNA extracted by CTAB were not clear, whereas the bands of RNA extracted by the TRIzol kit were brighter and clearer, indicating higher quality and integrity of the RNA products extracted by the TRIzol kit. The 28S rRNA band was approximately 1.5- to 2-fold brighter than the 18S rRNA band on the agarose gel electrophoresis. The absorbance value A 260/280 was 1.8-2.0, and the absorbance value A 260/230 was 〉1.9. The Bioanalyzer RNA integrity results showed that the RNA integrity number of the RNA extracted using the TR/zol kit was acceptable for high-throughputsequencing. Therefore, the total RNA extracted using the TRIzol kit method can be used for high-throughput sequencing and other molecular biology experiments.
文摘副溶血弧菌(Vibrio parahaemolyticus)是一种革兰氏阴性嗜盐菌。提取高质量的RNA是研究副溶血弧毒力基因表达与其致病性和环境适应性的基础。本研究分别用SDS法、Trizol法,PureLinkTM RNA Mini Kit和Uniq-10柱式Trizol总RNA抽提试剂盒提取六株副溶血弧菌的总RNA,用普通琼脂糖凝胶电泳和核酸测定仪检测RNA的质量,并用RT-PCR法对四种提取方法进行比较。研究结果表明,Trizol法和Pure-LinkTM RNA Mini Kit简单快捷,提取的总RNA完整性好,纯度高,可用于后续实验的分子生物学研究;而Trizol法更适用于普通实验室细菌RNA的提取。
文摘为建立草莓果实总RNA的提取方法,针对草莓成熟果实中富含多糖、多酚和色素等次级代谢物质,总RNA提取难度大的特点,比较改良的EASYspin植物RNA提取试剂盒法和改良的CTAB法提取草莓果实中总RNA的质量。通过琼脂糖凝胶电泳和核酸蛋白测定仪(Nano Drop 2000)检测2种方法提取总RNA的浓度、纯度及完整性等。改良的EASYspin植物RNA提取试剂盒法和改良的CTAB法都能够完成草莓果实总RNA的提取,电泳检测在28S和18S处呈2条清晰的条带,OD260/OD280值和OD260/OD230值均在2.0左右;改良的EASYspin植物RNA提取试剂盒法成本较高,且提取总RNA质量劣于改良的CTAB法,但是EASYspin植物RNA提取试剂盒法操作简便,安全可靠,节省时间。通过RT-PCR验证,2种方法所获得的草莓果实总RNA质量较好,可达到分子生物学试验对RNA质量的要求。
