AllPrep DNA/RNA/Protein Mini Kit

For simultaneous purification of DNA, RNA, and protein from cells and tissues

Features

  • Maximal recovery of each analyte from precious samples
  • Time savings through short, streamlined workflow
  • Comparable data as analytes come from same sample
  • No need for organic solvents such as phenol and acetone
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AllPrep DNA/RNA/Protein Mini Kit (50)

Cat. No. / ID: 80004

For 50 preps: AllPrep DNA Spin Columns, RNeasy Spin Columns, Collection Tubes, RNase-Free Reagents and Buffers
SEK 7,385.00
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The AllPrep DNA/RNA/Protein Mini Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Product Details

The AllPrep DNA/RNA/Protein Mini Kit allows simultaneous purification of DNA, RNA, and protein from the same cell or tissue sample. The kit can be used in combination with Allprotect Tissue Reagent, which delivers immediate stabilization of DNA, RNA, and protein in tissue samples for long-term storage without freezing. Tissues can be efficiently disrupted using a TissueRuptor or TissueLyser system.

Performance

Genomic DNA and total RNA purified using the AllPrep DNA/RNA/Protein Mini Kit deliver optimal results in real-time PCR and real-time RT-PCR analyses (see figure " Reliable real-time PCR and RT-PCR analyses"). Total protein purified using the kit is suitable for SDS-PAGE and western blotting (see figure " Reliable western blotting and SDS-PAGE").
See figures

Principle

The AllPrep DNA/RNA/Protein Mini Kit standardizes sample preparation for systems biology. DNA, RNA, and protein are all prepared from the same source, eliminating the variation inherent in preparing these analytes from different samples.There is no need to split the sample into 3 prior to purification, allowing maximum recovery of DNA, RNA, and protein. In addition, no organic solvents such as phenol and acetone are required.

Procedure

Cells or tissues are first lysed and homogenized in Buffer RLT, which immediately inactivates DNases and RNases as well as proteases to ensure isolation of intact DNA, RNA, and proteins. The lysate is then passed through an AllPrep DNA spin column. This column, in combination with the high-salt buffer, allows selective and efficient binding of genomic DNA. The column is washed and pure, ready-to-use DNA is then eluted.

Ethanol is added to the flow-through from the AllPrep DNA spin column to provide appropriate binding conditions for RNA, and the sample is then applied to an RNeasy spin column, where total RNA binds to the membrane and contaminants are efficiently washed away. High-quality RNA is then eluted in RNase-free water.

Buffer APP, a novel aqueous protein precipitation solution, is added to the flow-through of the RNeasy spin column, and the precipitated proteins are pelleted by centrifugation. Intact total proteins are redissolved in an appropriate buffer and then ready to use in downstream applications. The kit includes Buffer ALO, which is compatible with SDS-PAGE, for dissolving the protein pellet. (See flowchart " AllPrep DNA/RNA/Protein procedure".) 

See figures

Applications

The AllPrep DNA/RNA/Protein Mini Kit streamlines sample preparation by enabling simultaneous purification of genomic DNA, total RNA, and total protein from the same cell or tissue sample. Total protein purified using the kit is suitable for SDS-PAGE and western blotting. Genomic DNA and total RNA purified using the AllPrep DNA/RNA/Protein Mini Kit deliver optimal results in all downstream applications including:

  • Real-time PCR analysis
  • Real-time RT-PCR analysis

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsPCR, RT-PCR,real-time PCR, real-time RT-PCR, Southern-,Northern-,Western Blotting, Microarrays, Genotyping,1D and 2D gel electrophoresis
Main sample typeEasy-to-lyse tissue or cells
Elution volumeRNA: 30 - 50µl, DNA: 100 - 200µl
Purification of total RNA, miRNA, poly A+ mRNA, DNA or proteinTotal RNA, genomic DNA and western-grade proteins
FormatSpin column
Yieldvaries, depends on sample type
Time per run or per prep60 min
ProcessingManual (centrifugation)
TechnologySilica technology, Protein precipitation
Sample amountup to 30 mg tissue or 1 x 10e7 cells

Resources

Kit Handbooks (1)
For simultaneous purification of genomic DNA, total RNA, and total protein from the same cell or tissue sample
Quick-Start Protocols (1)
For simultaneous purification of genomic DNA, total RNA and total protein from the same cell or tissue sample
Brochures & Guides (2)

FAQ

Can the AllPrep DNA/RNA/Protein Mini Kit be used with fibrous or lipid tissues?

The AllPrep DNA/RNA/Protein Mini Kit is optimized for use with easy-to-lyse tissues. For fibrous (e.g., heart) and lipid tissues (e.g., brain), we recommend using a portion of the tissue sample for RNA purification with the RNeasy Fibrous Tissue Mini Kit and RNeasy Lipid Tissue Mini Kit, respectively.

 

 

 

FAQ ID -1579
What is the composition of Buffer ALO?
The detailed composition of Buffer ALO is confidential. Buffer ALO is part of the AllPrep DNA/RNA/ Protein Kit and functions as protein resuspension buffer. Buffer ALO is a Laemmli-related sample buffer for use in SDS-PAGE. In addition to other reagents, Buffer ALO comprises SDS, bromophenol blue, and dithiothreitol (DTT). DTT is added by the user prior the first use of the kit.
FAQ ID -2805
Does Buffer APP in the AllPrep DNA/RNA/Protein Mini Kit contain acetone?

No, Buffer APP in the AllPrep DNA/RNA/Protein Mini Kit is a novel aqueous protein precipitation solution and has some advantages over acetone with regard to recovery of small proteins (about 10 kDa and smaller).

 

 

FAQ ID -1576
What effect does homogenization have on DNA yield and integrity when using AllPrep DNA/RNA Kits?

Yield and integrity of genomic DNA depend on the disruption and homogenization method used with the Allprep DNA/RNA Kits.

Homogenization with the TissueRuptor (or other rotor–stator homogenizer, such as the Polytron) or the TissueLyser results in greater DNA fragmentation (depending on homogenization time and intensity). However, shorter DNA fragments are easier to elute, so DNA yields will be higher.

In contast, homogenization using the QIAshredder (or a syringe and needle) is more gentle, resulting in less DNA fragmentation. However, longer DNA fragments are more difficult to elute, so DNA yields may be lower.

 

FAQ ID -1751
What can be done if the protein pellet does not solubilize completely using the AllPrep DNA/RNA/Protein Mini Kit?

In the protein precipitation procedure of the AllPrep DNA/RNA/Protein protocol, be sure to dissolve the protein pellet completely in Buffer ALO or 1x SDS-PAGE sample buffer.

For easier dissolving of proteins, dissolve the pellet in 5% (w/v) SDS or 8 M urea. Commonly, there is insoluble material left even after vigorous shaking or heating. This is typical for all kinds of protein precipitation under heavily denaturing conditions.

Briefly centrifuge the sample and use the supernatant for downstream analysis.

 

FAQ ID -1580
What is the composition of Buffer APP?
The exact composition of Buffer APP is confidential. Buffer APP is a component of the AllPrep DNA/RNA Protein Kit and functions as a protein precipitation buffer, containing, among other components, zinc chloride.
FAQ ID -2804
Can acetone be used for precipitation of protein from Buffer RLT lysates generated with RNeasy Kits?

Yes, please follow the Supplementary Protocol Acetone precipitation of protein from Buffer RLT or Buffer RLT Plus lysates (RY22).

Important Note:

Do not use TCA to precipitate protein from Buffer RLT and Buffer RLT Plus lysates. These buffers contain guanidine thiocyanate, which can form highly reactive compounds when combined with acidic solutions.

 

For simultaneous purification of DNA, RNA, and protein from the same sample (either cultured cells or easy-to-lyse tissues), we recommend using the AllPrep DNA/RNA/Protein Mini Kit. This kit allows precipitation of protein from Buffer RLT lysates using a novel protein precipitation buffer, Buffer APP.

Please note that Buffer APP is not compatible with Buffer RLT Plus. Acetone should be used instead to precipitate protein from RLT Plus lysates.

 

FAQ ID -1164
Can tissue homogenized in Buffer RLT Plus instead of Buffer RLT be used with the AllPrep DNA/RNA/Protein Mini Kit?

No, Buffer APP for protein precipitation in the Allprep DNA/RNA/protein Mini Kit is not compatible with Buffer RLT Plus. Acetone can be used to precipitate proteins from samples homogenized in Buffer RLT Plus contained in the Allprep DNA/RNA Mini Kit and RNeasy Plus Mini Kit.  

 

FAQ ID -1577
Why does AllPrep DNA/RNA/Protein Mini kit use buffer RLT, but Allprep DNA/RNA Mini and Allprep DNA/RNA Micro kits use buffer RLT Plus? Are these buffers interchangeable?

Buffer RLT Plus contains detergents that are more ideal for binding of DNA to the AllPrep DNA mini spin columns.

But RLT Plus is not compatible with Buffer APP used to precipitate proteins in AllPrep DNA/RNA/Protein Mini kit. Therefore, we chose RLT instead of RLT Plus to get both DNA and proteins with this kit.

RLT lysates can also be used in AllPrep RNA/DNA kit or the AllPrep DNA/RNA Micro kit. DNA and RNA yields should be fine, but RNA may contain slightly higher levels of genomic DNA contamination.

FAQ ID - 3391
3323 - Do you have a protocol for the AllPrep DNA/RNA/Protein Mini Kit on the QIAcube?
A custom protocol can be requested through the online QIAcube protocol request page: <https://www.qiagen.com/qiacube/customized/protocoladd.aspx >.
FAQ ID - 3323