Product Features
- Consistent DNA binding and reproducibility across columns
- Fast and convenient, no toxic reagents needed
- No ethanol precipitation required
- High purity and quantity of plasmid DNA
- Suitable for enzyme digestion, transformation, PCR, in vitro transcription, and sequencing
Introduction
This kit utilizes a modified SDS-alkaline lysis method to lyse cells. The silica-based membrane in the spin column selectively binds plasmid DNA from the solution under high salt and low pH conditions. Impurities and other bacterial components are then removed using a deproteinization solution and a wash buffer. Finally, a low-salt, high-pH elution buffer is used to elute the purified plasmid DNA off the silica membrane.
Kit Components
- Equilibration Buffer
- RNase A
- Solution P1
- Solution P2
- Solution P3
- Protein Precipitation Solution
- Wash Buffer
- Elution Buffer
- Spin Column
- Collection Tubes
Assay Procedure Summary
- Prepare the spin column with equilibration buffer.
- Collect and pellet the bacterial cells by centrifugation.
- Resuspend the pellet in Solution P1.
- Lyse cells by adding Solution P2 and mixing gently.
- Neutralize with Solution P3, mix, and centrifuge.
- Transfer the supernatant to the spin column.
- Wash with Protein Precipitation Solution and discard liquid.
- Wash twice with Wash Buffer and discard liquid.
- Remove residual wash buffer by centrifugation.
- Elute plasmid DNA with Elution Buffer.
Storage/Stability
Store RNase A and Solution P1 at 4°C and rest of kit components at room temperature. Stable for 12 months if stored properly.