.png){kind=icon}
返回
A precise mRNA quantification method using CE-based SSCP
.png){kind=icon}
PRE
AI
DOI：10.1002/elps.200600225.png)
.png){kind=icon}
原文求助.png){kind=icon}
分享.png){kind=icon}
收藏
摘要
En 中文
Even though mRNA quantification provides significant information for biological analysis, current methods such as Northern blot analysis and real-time PCR are known to be laborious and lacking in precision. In this study, we demonstrate a new precise mRNA quantification method using CE based on SSCP (CE-SSCP) coupled with reverse transcription. mRNA samples could be simply analyzed for the quantification directly with reverse transcript obtained from a single reaction. This helps to avoid considerable errors generated by a series of the tedious manual steps. Also, unlike real-time PCR, reverse transcripts can be directly quantified by CE-SSCP in this method without further data estimation. Reproducibility and accuracy of CE-SSCP for mRNA quantification was examined using enhanced green fluorescent protein (eGFP) mRNA transcribed in vitro. Specific reverse transcription primer was determined for the accurate quantification of eGFP mRNA from total RNA obtained from the recombinant Escherichia coli. Using elongation factor TO mRNA as an internal standard, it was shown that sample-to-sample variation could be minimized. Expression kinetics at both mRNA level and protein level was studied and the potential of CE-SSCP in expression analysis was demonstrated by comparison with the eGFP activity assay.
Keyword:
CE
enhanced green fluorescent protein
expression analysis
mRNA quantification
SSCP
AI总结
对已上传原文的论文进行重点信息的提取，主要内容包括：简要概述、研究摘要、背景介绍、关键亮点、图文解析、展望与总结。
期刊
IF:
2.5
论文数:
1.2W
被引数:
1.0W
机构
暂无机构信息
引用论文
High-resolution quantification of specific mRNA levels in human brain autopsies and biopsies.png)
GENOME RESEARCH
OAAI
GENOME RESEARCHIF5.5
OAAI分享收藏A comparison of fluorescent SSCP and denaturing HPLC for high throughput mutation scanningHUMAN MUTATIONOAAI
HUMAN MUTATIONIF3.7
OAAI分享收藏Direct quantification of mRNA expression levels using single molecule detection使用单分子检测直接定量mRNA表达水平
PREAI
分享收藏分享收藏
查看更多内容