[Objective] The aim of this study was to compare three RNA extraction methods and thus find out the suitable one for isolating intact and high quality total RNA from Pinus bungeana.[Method] Employing three extraction ...[Objective] The aim of this study was to compare three RNA extraction methods and thus find out the suitable one for isolating intact and high quality total RNA from Pinus bungeana.[Method] Employing three extraction methods of Trizol,RNeasy Mini Kit,LiCl precipitation,total RNAs of P.bungeana were extracted from pine leaf samples,and their integrity and purity were detected via agarose gel electrophoresis and spectrophotometry for a comparative study.[Result] Among the three extraction methods,LiCl precipitation method demonstrated higher yield and better integrity of total extracted RNA,with OD260/OD280 ratio between 1.8-2.0 and clear 28 S and 18 S bands in electrophoresis pattern.[Conclusion] LiCl precipitation method could be used to extract highly pure and intact total RNA from P.bungeana.展开更多
The possible mechanism of inhalation anesthetics on the internal auditory impairment of the rat was investigated by determining the effect of nitrous oxide （N2O） and isoflurane on the total RNA yield from the cochle...The possible mechanism of inhalation anesthetics on the internal auditory impairment of the rat was investigated by determining the effect of nitrous oxide （N2O） and isoflurane on the total RNA yield from the cochlea of the rats. Thirty healthy Wistar rats were randomly divided into 3 groups： group C （control group, n=10） with a 3-h unremitting inhalation of 50% 02, group N （experiment group, n= 10） with a continuous inhalation of 50% N20＋50% O2for 3 h, and group I （experiment group, n=10） with a 3-h sustained inhalation of 2.5% isoflurane. The TRIzol in combination with RNeasy was used to respectively extract the total RNA from cochlea of rats in the 3 groups. Spectrophotometry was used to detect total RNA yield and electrophoresis to detect the quality. The total RNA extracted from the cochlea of the rats in the groups C and N was 7.69 and 6.51 μg, respectively. There was a 15% decrease in the N group as compared with group C. The total RNA from the rats in the group I was 7.32μg, and there was hardly any change in the group as compared with the group C. The value of A260/A280 in groups C, N and I was 2.07, 2.04 and 2.04, respectively, showing a very high RNA purity. The result of gel electrophoresis suggested that there was no degradation in the total RNA. It was suggested that the interference of N20 on the cochlear RNA yield might be one of the reasons which cause an injury of the ear. The isoflurane shows no harm on the hearing.展开更多
An effective method for isolation of total RNA from pulps of mainly deciduous fruit trees was discussed in this paper. Here we shown that total RNA was successfully isolated from pulps of apple, peach and strawberry, ...An effective method for isolation of total RNA from pulps of mainly deciduous fruit trees was discussed in this paper. Here we shown that total RNA was successfully isolated from pulps of apple, peach and strawberry, but not grape berry via the effective CTAB method modified by ZENG, et al. However, total RNA was isolated from grape berry according to ZENG＇s protocols with a modification including 16% of β-Mercatoethanol and 3% of PVP in extraction buffer, 1/3 （V/V） of 5mol/L potassium acetate （KAC PH4.8） in supernatant. The detection of the isolated total RNA above using ultraviolet spectrophoto-metry, electrophoresis, reverse transcription and RT-PCR shown that the CTAB and its modifications is a preferred method for isolation of high-quality total RNA from fruit pulps.展开更多
[ Objective] This study aimed to investigate the variation rule of total RNA concentration during formation of microtubers of PineUia ternata induced by sucrose. [ Method ] Plantlets of P. ternata were used as experim...[ Objective] This study aimed to investigate the variation rule of total RNA concentration during formation of microtubers of PineUia ternata induced by sucrose. [ Method ] Plantlets of P. ternata were used as experimental materials. Total RNA concentration was determined with spectrophotometer, and the variation of total RNA was analyzed before and after formation of P. ternata microtubers treated with different concentrations of sucrose using DDRT-PCR technology. [ Result] Total RNA concentration of P. ternata treated with different concentrations of sucrose varied greatly, which showed an upward trend in the first 15 d, de- clined from 15~ d to 25th d, and slightly raised from 25＇h d to 30＇8 d. Microtubers formed under various sucrose treatments showed an increase in RNA concentration after induction, and it was assumed that the treatment of high concentrations of sucrose might have activated the expression of development-related＇ genes in P. ter- nata microtubers. [ Conclusion] This study provides the basis for revealing the related gene expression information and molecular mechanism during formation of P. ternata microtubers induced by sucrose.展开更多
Leaves of Acer truncatum ＇ Luhong No. 1 ＇ contain large amounts of polysaccharides and polyphenols, which seriously affect extraction yield and quality of total RNA. In order to explore the appropriate total RNA ext...Leaves of Acer truncatum ＇ Luhong No. 1 ＇ contain large amounts of polysaccharides and polyphenols, which seriously affect extraction yield and quality of total RNA. In order to explore the appropriate total RNA extraction method, total RNA was extracted from leaves of A. truncatum ＇ Luhong No. 1 ＇ with three methods, including kit method, Trizol method and modified CTAB method. The results showed that ODE6o/OD2so and OD^o/OD2ao ratios of total RNA extracted from leaves of A. truncatum ＇ Luhong No. 1 ＇ with kit method were higher than 1.8, with a general yield and certain level of DNA contamination ; OD^o/OD~ and OD^o/OD^o ratios of total RNA extracted with Trizol method were about 1.5, with the lowest yield; OD260/OD280 and OD260/OD230 ratios of total RNA extracted with modified CTAB method were about 2.0, with the highest yield and distinct eleetrophoresis patterns. The results demonstrated that total RNA extracted with modified CTAB method exhibited high yield and purity, which could meet the demands of subsequent molecular biology research. Thus, modified CTAB method is the appro- oriate method for extracting total RNA from leaves of A. truncatum ＇ Luhong No. 1 ＇展开更多
探索一种新的适合草莓属(Fragariaspp.)组培苗、大田苗叶片总RNA的快速高效提取方法。试验以草莓叶片为材料,用改进的CTAB法对其RNA进行提取,并对提取的RNA进行了电泳检测、含量测定和RT-PCR检测。结果表明:用该RNA提取方法提取的RNA具...探索一种新的适合草莓属(Fragariaspp.)组培苗、大田苗叶片总RNA的快速高效提取方法。试验以草莓叶片为材料,用改进的CTAB法对其RNA进行提取,并对提取的RNA进行了电泳检测、含量测定和RT-PCR检测。结果表明:用该RNA提取方法提取的RNA具有28 S rRNA、18 S rRNA和5.8 S rRNA 3条清晰的条带,且无降解。OD260/OD280在1.83至2.14之间,具有较高的纯度,且含量较高。用该RNA提取方法提取的RNA逆转录成cDNA,经PCR扩增出现清晰的条带,说明该RNA提取方法提取的RNA可以满足进一步分子生物学试验的要求。展开更多
基金Supported by National Program on Key Basic Research Project(2009CB119204)~~
文摘[Objective] The aim of this study was to compare three RNA extraction methods and thus find out the suitable one for isolating intact and high quality total RNA from Pinus bungeana.[Method] Employing three extraction methods of Trizol,RNeasy Mini Kit,LiCl precipitation,total RNAs of P.bungeana were extracted from pine leaf samples,and their integrity and purity were detected via agarose gel electrophoresis and spectrophotometry for a comparative study.[Result] Among the three extraction methods,LiCl precipitation method demonstrated higher yield and better integrity of total extracted RNA,with OD260/OD280 ratio between 1.8-2.0 and clear 28 S and 18 S bands in electrophoresis pattern.[Conclusion] LiCl precipitation method could be used to extract highly pure and intact total RNA from P.bungeana.
基金The major financial support was from the Department of Education,Hubei Provincial Government (D200524008)was also the guidance project of the Scientific Foundation,Health Department,Hubei Provincial Government (JX2C32)
文摘The possible mechanism of inhalation anesthetics on the internal auditory impairment of the rat was investigated by determining the effect of nitrous oxide （N2O） and isoflurane on the total RNA yield from the cochlea of the rats. Thirty healthy Wistar rats were randomly divided into 3 groups： group C （control group, n=10） with a 3-h unremitting inhalation of 50% 02, group N （experiment group, n= 10） with a continuous inhalation of 50% N20＋50% O2for 3 h, and group I （experiment group, n=10） with a 3-h sustained inhalation of 2.5% isoflurane. The TRIzol in combination with RNeasy was used to respectively extract the total RNA from cochlea of rats in the 3 groups. Spectrophotometry was used to detect total RNA yield and electrophoresis to detect the quality. The total RNA extracted from the cochlea of the rats in the groups C and N was 7.69 and 6.51 μg, respectively. There was a 15% decrease in the N group as compared with group C. The total RNA from the rats in the group I was 7.32μg, and there was hardly any change in the group as compared with the group C. The value of A260/A280 in groups C, N and I was 2.07, 2.04 and 2.04, respectively, showing a very high RNA purity. The result of gel electrophoresis suggested that there was no degradation in the total RNA. It was suggested that the interference of N20 on the cochlear RNA yield might be one of the reasons which cause an injury of the ear. The isoflurane shows no harm on the hearing.
基金This work was supported by Beijing Natural Science Foundation （No. 6082005） and Funding Project for Academic Human Resources Development in Institutions of Higher Learning under the Jurisdiction of Beijing Municipality （No. PXM2007014207）.
文摘An effective method for isolation of total RNA from pulps of mainly deciduous fruit trees was discussed in this paper. Here we shown that total RNA was successfully isolated from pulps of apple, peach and strawberry, but not grape berry via the effective CTAB method modified by ZENG, et al. However, total RNA was isolated from grape berry according to ZENG＇s protocols with a modification including 16% of β-Mercatoethanol and 3% of PVP in extraction buffer, 1/3 （V/V） of 5mol/L potassium acetate （KAC PH4.8） in supernatant. The detection of the isolated total RNA above using ultraviolet spectrophoto-metry, electrophoresis, reverse transcription and RT-PCR shown that the CTAB and its modifications is a preferred method for isolation of high-quality total RNA from fruit pulps.
基金Supported by National Natural Science Foundation of China (30973963)Anhui Provincial Natural Science Foundation (090413252)Key Program of Natural Science Research of Anhui Provincial Universities (KJ2009A160)
文摘[ Objective] This study aimed to investigate the variation rule of total RNA concentration during formation of microtubers of PineUia ternata induced by sucrose. [ Method ] Plantlets of P. ternata were used as experimental materials. Total RNA concentration was determined with spectrophotometer, and the variation of total RNA was analyzed before and after formation of P. ternata microtubers treated with different concentrations of sucrose using DDRT-PCR technology. [ Result] Total RNA concentration of P. ternata treated with different concentrations of sucrose varied greatly, which showed an upward trend in the first 15 d, de- clined from 15~ d to 25th d, and slightly raised from 25＇h d to 30＇8 d. Microtubers formed under various sucrose treatments showed an increase in RNA concentration after induction, and it was assumed that the treatment of high concentrations of sucrose might have activated the expression of development-related＇ genes in P. ter- nata microtubers. [ Conclusion] This study provides the basis for revealing the related gene expression information and molecular mechanism during formation of P. ternata microtubers induced by sucrose.
基金Supported by Project of Agricultural Fine Varieties of Shandong Province[LKNZ(2012)No.213]
文摘Leaves of Acer truncatum ＇ Luhong No. 1 ＇ contain large amounts of polysaccharides and polyphenols, which seriously affect extraction yield and quality of total RNA. In order to explore the appropriate total RNA extraction method, total RNA was extracted from leaves of A. truncatum ＇ Luhong No. 1 ＇ with three methods, including kit method, Trizol method and modified CTAB method. The results showed that ODE6o/OD2so and OD^o/OD2ao ratios of total RNA extracted from leaves of A. truncatum ＇ Luhong No. 1 ＇ with kit method were higher than 1.8, with a general yield and certain level of DNA contamination ; OD^o/OD~ and OD^o/OD^o ratios of total RNA extracted with Trizol method were about 1.5, with the lowest yield; OD260/OD280 and OD260/OD230 ratios of total RNA extracted with modified CTAB method were about 2.0, with the highest yield and distinct eleetrophoresis patterns. The results demonstrated that total RNA extracted with modified CTAB method exhibited high yield and purity, which could meet the demands of subsequent molecular biology research. Thus, modified CTAB method is the appro- oriate method for extracting total RNA from leaves of A. truncatum ＇ Luhong No. 1 ＇
文摘探索一种新的适合草莓属(Fragariaspp.)组培苗、大田苗叶片总RNA的快速高效提取方法。试验以草莓叶片为材料,用改进的CTAB法对其RNA进行提取,并对提取的RNA进行了电泳检测、含量测定和RT-PCR检测。结果表明:用该RNA提取方法提取的RNA具有28 S rRNA、18 S rRNA和5.8 S rRNA 3条清晰的条带,且无降解。OD260/OD280在1.83至2.14之间,具有较高的纯度,且含量较高。用该RNA提取方法提取的RNA逆转录成cDNA,经PCR扩增出现清晰的条带,说明该RNA提取方法提取的RNA可以满足进一步分子生物学试验的要求。
