JoVE 비디오를 활용하시려면 도서관을 통한 기관 구독이 필요합니다. 전체 비디오를 보시려면 로그인하거나 무료 트라이얼을 시작하세요.
RIBO-seq in Bacteria: a Sample Collection and Library Preparation Protocol for NGS Sequencing
Chapters
- 00:00Introduciton
- 00:54Cell Harvesting and Lysis
- 04:13MNase Digestion and Size Selection of Samples for RIBO-seq
- 06:17rRNA Depletion and Alkaline Fragmentation of Samples for RNA-seq
- 07:07Library Preparation and Size Selection of cDNA Libraries
- 08:49Results: Quality of cDNA Libraries and Reads
- 11:31Conclusion
Here we describe the stages of sample collection and preparation for RIBO-seq in bacteria. Sequencing of the libraries prepared according to these guidelines results in sufficient data for comprehensive bioinformatic analysis. The protocol we present is simple, uses standard laboratory equipment and takes seven days from lysis to obtaining libraries.
