This is a free preview. For full access
Analyzing Gene Expression and Function
Ribosome Profiling for Translation Snapshot
Description
Ribosome profiling is a deep sequencing method that gives a snapshot of active translation in a cell. It sequences the mRNAs protected by ribosomes. This shows which messages are being read to make proteins at a specific moment.
The method has several uses in biology. It can track translation in ...
Show More
Transcript
确定基因组中被转录并翻译成蛋白质的区域,是理解基因组及其表达的关键步骤。
这一目标通过一种称为核糖体谱分析(ribosome profiling)的技术实现,该技术也被称为核糖体测序(ribo-seq)。该方法可定位核糖体在mRNA上的位置,并鉴定出正在被主动翻译成蛋白质的mRNA分子。
为了分离RNA,首先需要裂解细胞以获取其中的分子。随后用RNase处理裂解液,这些酶会降解未被核糖体覆盖的mRNA,仅留下受保护的mRNA片段。
核糖体保护的片段随后通过蔗糖梯度离心与未受保护的、被切割的片段分离。
随后,核糖体从 mRNA 片段上被移除,再通过逆转录酶进行 RT-PCR,将 RNA 转化为 DNA。
接下来,对 D...
Show More
Tags
More videos by Standards
Show More