RNA extraction principlesSummary: Principle and introduction of RNA isolation.
Content: High quality RNA isolation is a critical step for any downstream application such as northern blot, nuclease protection assay, RT-PCR, in vitro transcription, cDNA library construction, to name a few. Various factors may affect the quality of RNA extracted from tissue and cell samples.
1. Cell and tissue sample handling: It is important to inactivate endogenous RNases immediately after tissue or cell harvesting. Cell and tissue samples can be extracted immediately after harvesting in strong chaotropic lysis buffer. Or samples can be stored immediately in liquid nitrogen. Once freezed in liquid nitrogen, never let the sample thaw before RNA isolation. Samples can also be placed immediately into RNAlater solution available from Ambion.
2. Homogenize samples thoroughly to prevent RNA degradation and increase yields.
3. RNA storage conditions: RNA can be stored soluble in RNase-free water or as NH4OAc/ethanol precipitation and stored at -80c.
Keywords: RNA extraction
Last update: 2007-03-09 05:08:55 |
