AllPrep DNA/RNA/Protein Mini Kit (50)

Designed for simultaneous purification of DNA, RNA, and protein from cells and tissues, this QIAGEN kit streamlines sample preparation for systems biology. All three analytes are prepared from the same source, eliminating the variation inherent in preparing them from different samples. There is no need to split the sample into three prior to purification, allowing maximum recovery of DNA, RNA, and protein, and no organic solvents such as phenol and acetone are required.

Cells or tissues are first lysed and homogenized in Buffer RLT, which immediately inactivates DNases, RNases, and proteases to ensure isolation of intact DNA, RNA, and proteins. The lysate is then passed through an AllPrep DNA spin column, where genomic DNA selectively binds in combination with a high-salt buffer. After washing, pure ready-to-use DNA is eluted. Ethanol is added to the flow-through to provide appropriate binding conditions for RNA, which is then applied to an RNeasy spin column. Contaminants are efficiently washed away, and high-quality RNA is eluted in RNase-free water. Buffer APP, a novel aqueous protein precipitation solution, is then added to the flow-through, and the precipitated proteins are pelleted by centrifugation. The protein pellet is redissolved using Buffer ALO, which is compatible with SDS-PAGE.

The kit is optimized for use with easy-to-lyse tissues or cells, with a sample capacity of up to 30 mg of tissue or 1 x 10e7 cells. It can be used in combination with Allprotect Tissue Reagent for immediate stabilization of DNA, RNA, and protein in tissue samples for long-term storage without freezing, and tissues can be efficiently disrupted using a TissueRuptor or TissueLyser system. The kit uses silica technology and protein precipitation in a spin column format with manual processing by centrifugation. Elution volumes are 30-50 µl for RNA and 100-200 µl for DNA, and the procedure takes approximately 60 minutes per prep. Purified genomic DNA and total RNA are suitable for downstream applications including PCR, RT-PCR, real-time PCR, real-time RT-PCR, Southern, Northern, and Western blotting, microarrays, genotyping, and 1D and 2D gel electrophoresis, while total protein is suitable for SDS-PAGE and western blotting.