采用改良的Trizol法提取厚皮甜瓜‘银帝’果实中的RNA.利用琼脂糖凝胶电泳、紫外分光光度法和RT-PCR法进行纯度、浓度和RNA完整性检测.结果表明：1 mL Trizol用于提取200 mg甜瓜果实量时,得到的RNA产量最大,氯仿抽提2次时提取到的RNA的A...采用改良的Trizol法提取厚皮甜瓜‘银帝’果实中的RNA.利用琼脂糖凝胶电泳、紫外分光光度法和RT-PCR法进行纯度、浓度和RNA完整性检测.结果表明：1 mL Trizol用于提取200 mg甜瓜果实量时,得到的RNA产量最大,氯仿抽提2次时提取到的RNA的A260/A280在1.9-2.0之间,所提取的RNA无蛋白质和DNA污染,室温沉淀和低温沉淀对RNA的产量无明显影响.将提取的RNA反转录后扩增β-actin基因片段,扩增的条带与设计大小一致,验证了该方法提取的RNA可用于RT-PCR.展开更多
对采自湖北神农架的神农香菊(D end ranthem a ind icum(L.)D es M on l.var.arom a ticum Q.H.L iu et S.F.Zhang var.nov.)的化学成分进行研究,从中分离到5个化合物,利用波谱分析技术和文献对照确定了其结构,分别鉴定为木犀草素(lu te...对采自湖北神农架的神农香菊(D end ranthem a ind icum(L.)D es M on l.var.arom a ticum Q.H.L iu et S.F.Zhang var.nov.)的化学成分进行研究,从中分离到5个化合物,利用波谱分析技术和文献对照确定了其结构,分别鉴定为木犀草素(lu teo lin),木犀黄酮甙(lu teo lin-7-O--βg lucopyranos ide),刺槐素甙(acacetin-7-rham nos idg luos ide),1-单山艹俞酸甘油酯(g lycery l-1-m onobehenate),山艹俞酸(behen ic ac id)。展开更多
After many unsuccessful attempts to obtain biologically active mRNAs from dry seeds of Davidia involucrata using available protocols,an effective procedure modified from Vicient and Delseny (1999) is developed.This me...After many unsuccessful attempts to obtain biologically active mRNAs from dry seeds of Davidia involucrata using available protocols,an effective procedure modified from Vicient and Delseny (1999) is developed.This method is based on the use of 8mol/L LiCl in the extraction buffer followed by phenol extractions.The differences with respect to the original method are the inclusions of 2% soluble PVP (10000Da) and 10 mmol/L cysteine in the extraction buffer to avoid oxidation of polyphenolics,the reduction in the number of phenol extractions,freezing the seeds for 10 minutes instead of using quarts.The author modifications conquer the perishing oxidation of polyphenolics,in their oxidized forms,polyphenols can covalently bind to proteins and nucleic acids; and permit the recovery of and an average of 240 μg total RNA per g of the seeds that is suitable for translation in vitro,northern hybridizations,and the construction of cDNA libraries.展开更多
文摘采用改良的Trizol法提取厚皮甜瓜‘银帝’果实中的RNA.利用琼脂糖凝胶电泳、紫外分光光度法和RT-PCR法进行纯度、浓度和RNA完整性检测.结果表明：1 mL Trizol用于提取200 mg甜瓜果实量时,得到的RNA产量最大,氯仿抽提2次时提取到的RNA的A260/A280在1.9-2.0之间,所提取的RNA无蛋白质和DNA污染,室温沉淀和低温沉淀对RNA的产量无明显影响.将提取的RNA反转录后扩增β-actin基因片段,扩增的条带与设计大小一致,验证了该方法提取的RNA可用于RT-PCR.
文摘对采自湖北神农架的神农香菊(D end ranthem a ind icum(L.)D es M on l.var.arom a ticum Q.H.L iu et S.F.Zhang var.nov.)的化学成分进行研究,从中分离到5个化合物,利用波谱分析技术和文献对照确定了其结构,分别鉴定为木犀草素(lu teo lin),木犀黄酮甙(lu teo lin-7-O--βg lucopyranos ide),刺槐素甙(acacetin-7-rham nos idg luos ide),1-单山艹俞酸甘油酯(g lycery l-1-m onobehenate),山艹俞酸(behen ic ac id)。
文摘After many unsuccessful attempts to obtain biologically active mRNAs from dry seeds of Davidia involucrata using available protocols,an effective procedure modified from Vicient and Delseny (1999) is developed.This method is based on the use of 8mol/L LiCl in the extraction buffer followed by phenol extractions.The differences with respect to the original method are the inclusions of 2% soluble PVP (10000Da) and 10 mmol/L cysteine in the extraction buffer to avoid oxidation of polyphenolics,the reduction in the number of phenol extractions,freezing the seeds for 10 minutes instead of using quarts.The author modifications conquer the perishing oxidation of polyphenolics,in their oxidized forms,polyphenols can covalently bind to proteins and nucleic acids; and permit the recovery of and an average of 240 μg total RNA per g of the seeds that is suitable for translation in vitro,northern hybridizations,and the construction of cDNA libraries.
