摘要
荔枝果肉因含糖量高、水分大和含较高活性的氧化酶而难以提取高质量的总RNA。以妃子笑荔枝不同发育期果肉为试材,经过多种提取方法比较和改进,发现市售RNA提取试剂盒改进提取步骤后均可快速得到质量较高的总RNA,果肉总RNA完整性好、质量高,平均浓度都在20μg/g以上,符合进一步分子生物学试验的要求。以反转录后得到的cDNA为模板,进一步的实时定量PCR反应表明以未纯化的RNA反转录的cDNA为模板没有信号,而经纯化后即可得到有规律的基因表达。
To isolate high quality total RNA from litchi aril is difficult because the tissue contain high levels of sugars, water and oxidase. In this study, total RNA of ＇FZX＇ litchi aril on different sampling dates was extracted with some methods and obtained high quality total RNA extracted with the improved extraction kit. The total RNA was indicated to be perfectly intact and good quality tested by Nucleic acid/protein assay and agarose gel electrophoresis. The yields of extracted total RNA were all more than 20 μg/g fresh weight and were suitable for further molecular biology analysis, such as Real-Time PCR etc. The results of Real-Time PCR showed that there displayed a regularly marked signal on the gene expression using the cDNA as templates which were synthesized from the total RNA extraction.
出处
《广东农业科学》
CAS
CSCD
北大核心
2012年第15期166-168,175,共4页
Guangdong Agricultural Sciences
基金
国家自然科学基金(30971985)
国家公益性行业(农业)科研专项(200903044)
国家现代农业产业技术体系建设专项资金(CARS-33)
