Product Features
- High-quality silica membrane ensures consistent RNA binding
- No ethanol precipitation required
- Fast and simple; process sample in 30 minutes
- Suitable for RNAi, RT-PCR, Northern blot
Introduction
Recent extensive research on RNA interference and regulatory small RNAs has created a pressing need for a kit that can effectively extract RNA of approximately 15-30 nucleotides (including siRNA and miRNA). However, traditional RNA extraction methods, such as silica membrane, cannot effectively adsorb and recover small RNA. Phenol/guanidine extraction and ethanol precipitation cannot effectively precipitate and recover small RNA. This kit uses a unique lysis buffer/β-mercaptoethanol to rapidly lyse cells and inactivate RNA-degrading enzymes. Strong organic extraction removes proteins and DNA. RNA, including small RNA, is adsorbed onto a special silica membrane in the spin column. A series of rapid wash-centrifugation steps further removes impurities such as cellular metabolites and proteins. Finally, a low-salt elution buffer elutes pure RNA from the silica membrane.
Kit Components
- Lysis/Binding Buffer
- 70% Alcohol
- Wash Solutions
- RNase-free H2O
- Spin Columns and Collection Tubes
Assay Procedure Summary
- Collect cells or tissue and add lysis buffer to break them down.
- Mix thoroughly to ensure complete lysis and inactivation of RNase.
- Add chloroform, mix, and separate the phases by centrifugation.
- Transfer the upper phase to a new tube and add ethanol.
- Load the mixture onto a spin column and centrifuge.
- Wash the column with Wash Solutions.
- Centrifuge to remove residual wash buffer.
- Elute RNA with RNase-free water.
- Use the purified RNA for downstream applications.
Storage/Stability
Store Lysis/Binding Buffer at 4°C and rest of kit components at room temperature. Stable for 6 months if stored properly.