mRNA isolation (Micro-FaskTrack kit)
- 최초 등록일
- 2014.09.13
- 최종 저작일
- 2014.08
- 3페이지/ 한컴오피스
- 가격 1,000원
목차
1. Introduction
2. Overview
3. Materials
4. Method
5. References
본문내용
★ The Micro-FastTrack™2.0 mRNA Isolation Kit allows isolation of polyA+ RNA directly from cells (1 x 102 - 5 x 106), tissue (10-200 mg), or total RNA (100-500 µg) in 2-3 hours using minimal equipment (a water bath, a microcentrifuge, and a syringe fit with an 18-21 gauge needle) without the need for ultracentrifugation or guanidinium lysis.
★ A typical mammalian cell contains about 10-11 g of RNA of which 1-5% is polyA+ RNA. The remaining RNA is mostly rRNA (80-85% of total) and low-molecular weight RNAs such as tRNA (15-20% of total). To separate the heterogeneous population of mRNA from the majority of the RNA found in the cell, affinity binding to oligo(dT) cellulose is used. This method exploits the major characteristic of mRNA, polyadenylation, to obtain intact, pure mRNA.
<중 략>
1. Incubate the cell lysate produced in Preparing Samples at 45°C for 15-20 minutes.
Note: Incubation is important for complete digestion of proteins and ribonucleases. Incubation for 20 minutes is recommended for tissues, while cell material is effectively digested with a 15 minute incubation. You may optimize the time of incubation for your particular sample.
참고 자료
http://www.invitrogen.com. Instruction Manual. Micro-FastTrack. 2.0 Kit
Sambrook and Russell. 2001. Molecular cloning 3rd. 7.1-7.20