特点

•     · High-quality DNA and RNA from the same sample
•     · Maximal yields of DNA and RNA from precious samples
•     · Rapid purification with short, streamlined protocol
•     · Streamlined protocols for 96-well plates
•     · Ready-to-use DNA and RNA for any downstream analysis

产品详情

AllPrep DNA/RNA Kits enable simultaneous purification of genomic DNA and total RNA from each cell or tissue sample. Since there is no need to divide each sample into two for separate purification procedures, maximum yields of DNA and RNA can be achieved. The purified DNA and RNA are eluted separately.

The AllPrep DNA/RNA Micro and Mini Kits can be automated on the QIAcube Connect. The AllPrep DNA/RNA 96 Kit is well suited for high-throughput purification of genomic DNA and total RNA from up to 96 samples.

性能

Small samples of cultured cells and easy-to-lyse tissues can be processed with AllPrep DNA/RNA technology (see table “Optimal yields of DNA and RNA from tissues using the AllPrep DNA/RNA Micro Kit”). Genomic DNA is purified using the novel AllPrep DNA spin column, and total RNA is purified using an RNeasy MinElute spin column. Total RNA with Agilent RIN values of close to 10 is routinely obtained from cultured cells. Genomic DNA and total RNA show reliable quantification over a wide dynamic range (see figure " Wide dynamic range").

Optimal yields of DNA and RNA from tissues using the AllPrep DNA/RNA Micro Kit

* DNA and RNA were purified from two 5 mg samples of each tissue, and the average yields are shown.

A simple workflow allows DNA and RNA extraction from one sample. The purified genomic DNA has an average length of 15–30 kb and performs well in multiplex PCR (see figure  "Multiplex PCR of 8 targets"). Total RNA is of high quality and has a RIN value of 10 indicating that the RNA is intact (see figure " Simultaneous purification of total RNA and genomic DNA" and figure " High-quality total RNA").

AllPrep DNA/RNA technology also enables efficient purification of high-quality DNA and RNA from different cell and tissue types in convenient 96-well format (see table “High-quality RNA from tissues purified using the AllPrep DNA/RNA 96 Kit” and figure " Reproducible purification of DNA and RNA in 96-well format").

High-quality RNA from tissues purified using the AllPrep DNA/RNA 96 Kit

* Intact RNA is indicated by a RIN value of close to 10 analyzed on the Agilent Bioanalyzer 2100.

There is no need for additional RNase or DNase digestion (see figure " RNA with no genomic DNA contamination") and the purified DNA is suitable for downstream applications (see figure " High-quality DNA for reliable multiplex PCR analysis").

原理

AllPrep DNA/RNA Kits are designed to purify both genomic DNA and total RNA from the same cell or tissue sample. Since there is no need to divide the sample into two for separate purification procedures, maximum yields of DNA and RNA can be achieved. Efficient purification of high-quality DNA and RNA from different tissue types is achieved without the need for additional RNase or DNase digestion. The purified DNA and RNA are eluted separately and ready to use in any downstream application. Processing in 96-well format makes AllPrep DNA/RNA technology the ideal tool for sample preparation in genomics and systems biology.

步骤

A simple spin-column workflow allows the purification of high-quality DNA and RNA from the same sample (see flowchart “ AllPrep DNA/RNA procedure”). Both cultured cells and easy-to-lyse tissues can be processed. Genomic DNA is purified using the novel AllPrep DNA spin column, and total RNA is purified using an RNeasy Mini or RNeasy MinElute spin column (AllPrep DNA/RNA Micro Kit).

When disrupting and homogenizing tissues in Buffer RLT Plus (supplied with the AllPrep DNA/RNA Micro Kit and Allprep DNA/RNA Mini Kit), excessive foaming may occur (please note this is not the case for the Allprep DNA/RNA 96 Kit since this product is supplied with Buffer RLT). This foaming is substantially reduced by adding Reagent DX (supplied separately) to Buffer RLT Plus at a final concentration of 0.5% (v/v) before starting disruption and homogenization. Reagent DX has been carefully tested with the kits, and has no effect on RNA purity or on downstream applications such as real-time RT-PCR. Buffer RLT Plus containing Reagent DX can be stored at room temperature (15–25ºC) for at least 9 months.

A supplementary protocol allows optional purification of protein. For experiments where DNA, RNA, and protein are regularly prepared from each sample, the AllPrep DNA/RNA/Protein Mini Kit can be used instead.

For high-throughput processing, the 96-well purification plates of the AllPrep DNA/RNA 96 Kit are for both DNA and RNA. The plates are rapidly and conveniently processed using either a centrifuge (Centrifuge 4-16 and Plate Rotor 2 x 96) or a combination of vacuum (QIAvac 96) and centrifuge. Both cultured cells and easy-to-lyse tissues can be processed.

AllPrep DNA/RNA Kits form part of QIAGEN's solution for preparing multiple analytes from the same sample. These include Allprotect Tissue Reagent, which stabilizes DNA, RNA, and protein in tissues, and RNAprotect Cell Reagent, which stabilizes DNA and RNA in cells. Both reagents deliver immediate stabilization at room temperature. For tissues, TissueRuptor and TissueLyser systems provide fast disruption at low- to high-throughputs.

应用

The purified genomic DNA has an average length of 15–30 kb, depending on homogenization conditions, and is suited for any application, including next-generation sequencing, Southern-, dot-, and slot-blot analyses; and PCR and multiplex PCR.

The purified total RNA can be used in any application, such as RNA-seq RT-PCR and real-time RT-PCR; differential display; cDNA synthesis; northern-, dot-, and slot-blot analyses; and microarrays.

A supplementary protocol allows optional purification of protein. The purified protein is denatured and can be used in applications such as SDS-PAGE, western blotting, and 2D gel electrophoresis.

Comparison of AllPrep DNA/RNA Kits