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RNeasy Plus Kits

For fast purification total RNA from cells and tissues using gDNA Eliminator columns or plates

Features

  • Efficient gDNA removal with unique gDNA Eliminator columns or plates (no need for DNase)
  • High-quality total RNA in minutes using fast and simple extraction protocols
  • Phenol-free RNA isolation
  • High-throughput processing in 96-well format
  • Ideal for sensitive applications such as real-time RT-PCR and RNA-seq
RNeasy Plus Micro Kit (50)

Cat. No. / ID: 74034

For 50 micropreps: RNeasy MinElute Spin Columns, gDNA Eliminator Spin Columns, Collection Tubes, Carrier RNA, RNase-Free Water and Buffers
Column typePlate type
Micro
Mini
96 well
RNeasy Plus Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

Product Details

RNeasy Plus Kits are next generation QIAGEN RNA extraction kits, combining high-quality RNA purification with effective gDNA removal with unique gDNA Eliminator columns or plates. Total RNA of high purity and yield is obtained from a wide range of cultured cells and easy-to-lyse tissues. The RNeasy Plus Micro Kit is specially designed for limited amounts of samples and isolates up to 45 µg pure total RNA The RNeasy Plus Mini Kit purifies up to 100 µg total RNA (>200 nt) from a single extraction with efficient gDNA removal. Both kits can be automated on the QIAcube Connect and samples can be conveniently stabilized using RNAprotect Tissue Reagent or Allprotect Tissue Reagent and efficiently disrupted using a TissueRuptor II or TissueLyser II or LT system (available separately). The RNeasy Plus 96 Kit is for high-throughput purification of total RNA from cultured cells grown in 96-well plates. For optimal results it is recommended to use this product together with the QIAvac 96 System.
Request a quote for a trial RNeasy Plus Mini Kit.

RNeasy Plus Mini standard protocols can also be executed using the TRACKMAN Connected system, paired with PIPETMAN M Connected pipettes, both from Gilson. The TRACKMAN Connected system guides researchers through the RNeasy Plus Mini protocols while automatically adjusting the Bluetooth-enabled PIPETMAN M Connected pipette settings. Each step of the protocol execution is recorded to accelerate reporting by generating a comprehensive run report. Download more information.

Performance

RNA extraction from cells and tissues with RNeasy Plus Micro and Mini Kits allows high, reproducible RNA yields and efficient genomic DNA elimination for sensitive applications (see figures “ Effective cell genomic DNA removal”, “ Effective tissue genomic DNA removal”, " High, reproducible RNA yields", and “ Array-ready RNA”). Total RNA with Agilent RIN values of close to 10 is routinely obtained from tissues and cultured cells.


RNeasy Plus Micro and Mini Kits also provide an enrichment for mRNA since most RNAs <200 nucleotides (such as rRNAs and tRNAs) are excluded. For isolation of total RNA containing small RNAs (e.g., miRNA) from cultured cells, additional RNA isolation protocols are available and described in the RNeasy Plus Micro Handbook and as a separate Supplementary Protocol to the RNeasy Plus Mini Kit.
The RNeasy Plus 96 Kit is compatible with a wide range of cell lines, providing purification of high-quality RNA with Agilent RIN values of close to 10 (see figure " High-quality RNA"). Complete genomic DNA removal by the kit (see figure "Effective genomic DNA removal") enables specific transcript detection in highly sensitive applications, such as quantitative, real-time RT-PCR analysis of low-abundance targets. In addition, reliable RNA purification from 102 to 106 cells allows real-time RT-PCR quantification over a wide dynamic range (see figure " Reliable RNA purification").

See figures

Principle

The RNeasy Plus procedure integrates QIAGEN’s innovative technology for selective binding of double-stranded DNA with well-established RNeasy technology. Efficient purification of high-quality RNA is guaranteed, without the need for additional DNase digestion. The purified RNA is ready to use and is ideally suited for downstream applications that are sensitive to low amounts of DNA contamination, such as quantitative, real-time RT-PCR.


Cells and easy-to-lyse tissues are first lysed and homogenized in highly denaturing guanidine-isothiocyanate–containing Buffer RLT Plus, which immediately inactivates RNases to ensure isolation of intact RNA. The lysate is then passed through a gDNA Eliminator spin column or gDNA Eliminator 96 plate that, in combination with the high-salt buffer, selectively and efficiently removes genomic DNA. Ethanol is added to provide appropriate binding conditions for RNA, and the sample is applied to an RNeasy spin column or 96-well plate. These specialized columns and 96-well plates contain a silica membrane that specifically binds RNA from lysed cells.
For fatty and difficult-to-lyse tissues, RNeasy Plus Universal Kits are recommended. For isolation of total RNA including miRNA from other sample types, we recommend miRNeasy Kits.

Procedure


The RNeasy Plus Micro Kit purifies total RNA from up to 5 x 105 cells or 5 mg tissue, and the RNeasy Plus Mini Kit isolates total RNA from up to107 cells or 30 mg tissue. A short workflow enables RNA isolation with genomic DNA removal in less than 25 minutes (see flowchart  RNeasy Plus procedure). Samples are first lysed and homogenized. The lysate is passed through a gDNA Eliminator spin column, ethanol is added to the flow-through, and the sample is applied to an RNeasy spin column. RNA binds to the membrane and contaminants are washed away. High-quality RNA is eluted in as little as 14 µl water using the RNeasy Plus Micro Kit or 30 µl water using the RNeasy Plus Mini Kit.


Different protocols are available for different starting materials. The protocols differ mainly in the lysis and homogenization of the sample. Once the sample is applied to the gDNA Eliminator spin column, the protocols are similar. The procedure provides an enrichment for mRNA since most RNAs <200 nucleotides (such as rRNAs and tRNAs) are excluded.


When disrupting and homogenizing tissues in Buffer RLT Plus (supplied with the RNeasy Plus Mini Kit), excessive foaming may occur. This foaming is substantially reduced by adding Reagent DX (supplied separately) to Buffer RLT Plus before starting disruption and homogenization. Reagent DX has been carefully tested with the kit, and has no effect on RNA purity or on downstream applications.
High-throughput RNA purification is achieved through the use of RNeasy 96 plates. Prior to application to the RNeasy 96 plate, cell lysates are first passed through a gDNA Eliminator 96 plate, (see figure " RNeasy Plus 96 procedure"). The gDNA Eliminator 96 plates are conveniently processed using a centrifuge (Centrifuge 4-16). RNA purification using RNeasy 96 plates is manual, and comprises 3 simple steps: bind, wash, and elute. 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. Up to 2 x 106 cells can be used as starting material when processing plates using a centrifuge, while up to 1 x 106 cells can be used when processing plates using a vacuum and centrifuge. The procedure provides an enrichment for mRNA since most RNAs <200 nucleotides (such as rRNAs and tRNAs) are excluded. The RNeasy Plus procedure can be modified to allow the purification of total RNA containing small RNAs, such as miRNA, from cultured cells.

See figures

Applications

RNA purified using RNeasy Plus Kits is ideally suited for downstream applications that are sensitive to low amounts of DNA contamination, such as RNA-seq and quantitative real-time RT-PCR. The purified RNA can also be used in other applications.


The RNeasy Plus Micro Kit is suitable for small samples, including:

  • Laser-microdissected cryosections
  • Fine-needle aspirates

The ease with which RNA is purified in 96-well format makes the RNeasy Plus 96 Kit well suited for high-throughput gene expression analysis in areas such as:

  • Drug discovery
  • Biomedical research

Comparison of RNeasy Plus Kits

Features RNeasy Plus Micro Kit RNeasy Plus Mini Kit RNeasy Plus 96 Kit
Applications

RNA-seq, quantitative real-time RT-PCR, end-point RT-PCR,

Northern dot and slot blotting, microarray analysis

RNA-seq, quantitative real-time RT-PCR, end-point RT-PCR,

Northern dot and slot blotting, microarray analysis

RNA-seq, quantitative real-time RT-PCR, end-point RT-PCR,

Northern dot and slot blotting, microarray analysis

Elution volume 14 µl 30–50 µl 45–140 µl
Format Spin column Spin column 96-well plate
Integrated removal of genomic DNA Yes Yes Yes
Main sample type Tissue, cells Tissue, cells Cultured cells
Processing Manual (centrifugation) Manual (centrifugation) Manual (centrifugation, vacuum)

Purification of total RNA, miRNA,

poly A+ mRNA, DNA or protein

Total RNA Total RNA Total RNA
Sample amount 5 x 105 cells or 5 mg tissue 107 cells or 30 mg tissue 10 to 5 x 105 cells
Technology Silica technology Silica technology Silica technology
Time per run or per prep 25 minutes 25 minutes
Yield Varies Varies 1.3–3.1 µg (from 1 x 105 cells)

Supporting data and figures

Resources

Kit Handbooks (3)
Quick-Start Protocols (3)
Scientific Posters (1)
Poster for download
Supplementary Protocols (4)
There are two protocols: follow Protocol 1 if you want to purify total RNA containing miRNA, or follow Protocol 2 if you want to purify small RNA (includes miRNA, 5S rRNA, and tRNA) and larger RNA (>200 nt) separately.
Technical Information (1)

Publications

Caspase-2 involvement during ionizing radiation-induced oocyte death in the mouse ovary.
Hanoux V; Pairault C; Bakalska M; Habert R; Livera G;
Cell Death Differ; 2006; 14 (4):671-81 2006 Nov 3 PMID:17082817
Determinants of RNA quality from FFPE samples.
von Ahlfen S; Missel A; Bendrat K; Schlumpberger M;
PLoS One; 2007; 2 (12):e1261 2007 Dec 5 PMID:18060057
Insidious adrenocortical insufficiency underlies neuroendocrine dysregulation in TIF-2 deficient mice.
Patchev AV; Fischer D; Wolf SS; Herkenham M; Götz F; Gehin M; Chambon P; Patchev VK; Almeida OF;
FASEB J; 2006; 21 (1):231-8 2006 Nov 29 PMID:17135362
FGF19 is a target for FOXC1 regulation in ciliary body-derived cells.
Tamimi Y; Skarie JM; Footz T; Berry FB; Link BA; Walter MA;
Hum Mol Genet; 2006; 15 (21):3229-40 2006 Sep 25 PMID:17000708

FAQ

Can I buy the gDNA eliminator plates supplied in your RNeasy Plus 96 kit separately?

We do not sell the gDNA eliminator plates separately.

 

FAQ ID - 3389
What sample types can be used with the RNeasy Plus 96 Kit?

The RNeasy Plus 96 Kit provides high-throughput purification of total RNA from cultured cells grown in 96-well plates. There are protocols for small amounts of easy-to-lyse tissues, allowing RNA purification without the use of phenol using spin technology or vacuum/spin technology.  However, for most applications, we recommend the RNeasy 96 Universal Tissue Kit, which uses phenol-based sample lysis to enable RNA purification from all types of tissue.

 

FAQ ID -1991
Do you have a protocol for purification of miRNA from animal cells using the RNeasy Plus Mini Kit and RNeasy MinElute Cleanup Kit?
I received a kit containing the MinElute columns; however, they were left out for a while and not stored at 2–8°C upon receipt. Can I still use them?

The MinElute spin columns included in the following kits should be stored at 2–8°C upon arrival: AllPrep DNA/RNA Micro, EpiTect Fast DNA Bisulfite, EpiTect Fast FFPE Bisulfite, EpiTect Fast LyseAll Bisulfite, EpiTect Plus DNA Bisulfite, EpiTect Plus FFPE Bisulfite, EpiTect Plus LyseAll Bisulfite, exoRNeasy Serum/plasma Maxi, exoRNeasy Serum/Plasma Midi, GeneRead DNA FFPE, GeneRead rRNA Depletion, GeneRead Size Selection, MinElute Gel Extraction, MinElute PCR Purification, MinElute Reaction Cleanup, miRNeasy FFPE, miRNeasy Micro, miRNeasy Serum/Plasma, QIAamp DNA FFPE, QIAamp DNA Investigator, QIAamp DNA Micro, QIAamp MinElute Media, QIAamp MinElute Virus Spin, QIAamp MinElute Virus Vacuum, RNeasy FFPE, RNeasy Micro, RNeasy Plus Micro.

Short-term storage (up to 4 weeks) at room temperature (15–25°C) does not affect the performance. However, for optimal performance and quality, storage temperature should not exceed 25°C.

FAQ ID - 3560
Do you have a kit for RNA isolation from any kind of sample type?

The RNeasy 96 Universal Tissue Kit enables high-throughput purification of RNA from any animal or human tissue sample, including difficult-to-lyse fibrous and fatty tissues. For single tube format RNA purification, the RNeasy Plus Universal Kit is also available. 

Please refer to the Selection guide for RNA isolation for all sample types to find the optimal solution for your sample source.

FAQ ID -627
What is the composition of Buffer RLT?

The exact composition of Buffer RLT is confidential. This buffer is a proprietary component of RNeasy Kits. Buffer RLT contains a high concentration of guanidine isothiocycanate, which supports the binding of RNA to the silica membrane. Buffer RLT can be purchased separately (cat. no. 79216)

Note: note that ß-mercaptoethanol should be added to Buffer RLT before use to effectively inactivate RNAses in the lysate (10 µl ß-Mercaptoethanol per 1 ml Buffer RLT).

FAQ ID -2793
How can I check the integrity of RNA purified using RNeasy Kits?

The integrity and size distribution of total RNA purified with RNeasy Kits can be checked by denaturing-agarose gel electrophoresis, the Agilent 2100 bioanalyzer, or the QIAxcel Advanced System with the QIAxcel RNA QC Kit v2.0.

 

The respective ribosomal species should appear as sharp bands on the stained gel. 28S ribosomal RNA bands should be present with an intensity approximately twice that of the 18S RNA band. If the ribosomal bands are not sharp, but appear as a smear of smaller sized RNAs, it is likely that the RNA sample has suffered major degradation during preparation.

 

Size of ribosomal RNAs from various sources

 

Source rRNA Size (kb) E. coli 16S 1.5   23S 2.9 S. cerevisiae 18S 2.0   26S 3.8 Mouse 18S 1.9   28S 4.7 Human 18S 1.9   28S 5.0

 

 

 

 

 

 

 

 

 

FAQ ID -1024
Why is there no vacuum-only protocol for the RNeasy Plus 96 Kit?

During the gDNA elimination step of the RNeasy Plus 96 procedure, the flow-through of the gDNA Eliminator 96 plate has to be collected. If the plate were to be processed in a vacuum manifold, there would be a substantial risk of cross-contamination due to foaming of the lysate. Also, plate drying and RNA elution are more efficient with a centrifuge than with a vacuum manifold.

 

FAQ ID -1993
How much sample can be used with the RNeasy Plus 96 Kit?

The RNeasy Plus 96 Kit allows extraction of RNA from up to 1 x 10e6 cells with the vacuum/spin protocol, or up to 2 x 10e6 cells with the spin protocol.

 

 

FAQ ID -1992
How is the RNeasy Plus Universal Kit different from the RNeasy Plus Kit?

The RNeasy Plus Universal Mini Kit has been designed for optimal lysis of all types of tissue. Lysis and homogenization are performed using QIAzol Lysis Reagent, a phenol-guanidine based lysis reagent. The RNeasy Plus Mini Kit is the optimal solution for cells and easy-to-lyse tissue since lysis occurs with Buffer RLT. Additionally, the gDNA removal feature is based on a different technology. While the classic RNeasy Plus Kit contains gDNA eliminator columns, the RNeasy Plus Universal Kit contains a gDNA eliminator solution.

FAQ ID -2340
What is the maximum volume of RNA in solution that can be used with the QuantiTect Whole Transcriptome Kit?

A maximum of 5 µl RNA eluate from RNeasy extraction procedures can be added to the reverse-transcription reaction with the QuantiTect Whole Transcriptome Kit.

 

 

FAQ ID -1616
Can the protocols for purification of small RNAs from cells in the appendices of the RNeasy Plus 96 Handbook also be used with tissue samples?

No. Tissue lysis generates more debris than cell lysis. This additional debris is not compatible with the special binding conditions for small RNAs using the RNeasy Plus 96 Kit, and can lead to a reduction of up to 50% or more in RNA yield.

 

 

FAQ ID -1995
What is the difference between Buffers RLT and RLT Plus?

In comparison to Buffer RLT of, e.g., the RNeasy Mini Kit, Buffer RLT Plus of the RNeasy Plus Mini Kit and RNeasy Plus 96 Kit also contains a proprietary blend of detergents that aid in the binding of genomic DNA to the gDNA Eliminator Mini Spin Columns, or to the gDNA Eliminator 96 plate respectively.

FAQ ID -1043
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
Do I have to discard Buffer RLT with beta-Mercaptoethanol (ß-ME) added to it after 1 month of storage?
No. Beta-Mercaptoethanol (ß-ME) is stable for 1 month, but Buffer RLT itself is stable for at least 9 months at room temperature (15 to 25°C). Simply add fresh ß-ME to the Buffer RLT supplied in RNeasy Kits to ensure complete inactivation of RNases while isolating RNA.
FAQ ID -1037
How can foaming of Buffer RLT Plus lysates be avoided when using RNeasy Plus Micro and Mini Kits, or the Allprep DNA/RNA Mini Kit?

When using the RNeasy Plus Micro and RNeasy Plus Mini Kits, or the Allprep DNA/RNA Mini Kit, foaming of lysates can be substantially reduced by adding Reagent DX to Buffer RLT Plus at a final concentration of 0.5% (v/v) before starting sample lysis.

Reagent DX has been carefully tested with RNeasy Plus and Allprep Kits and has no effect on RNA purity or on downstream applications. Reagent DX can be purchased separately using catalog number 19088.

For more information on efficient sample disruption and homogenization for nucleic acid extractions, please see further product details and resources for the TissueRuptor, TissueLyser LT, and TissueLyser II.

FAQ ID -1734
What can be used as an alternative to the A260 measurement for quantification of small amounts of RNA and DNA?

Small amounts of RNA and DNA may be difficult to measure spectrophotometrically. Fluorometric measurements, or quantitative RT-PCR and PCR are more sensitive and accurate methods to quantify low amounts of RNA or DNA.

Fluorometric measurements are carried out using nucleic acid binding dyes, such as RiboGreen® RNA Quantitation Reagent for RNA, and PicoGreen® DNA Quantitation Reagent for DNA (Molecular Probes, Inc.).

FAQ ID -728
How efficient is DNA removal by a gDNA Eliminator 96 plate compared with DNase digestion on an RNeasy 96 plate?

Both DNase digestion on the RNeasy 96 plate as well as the gDNA Eliminator 96 plate with RNeasy Plus 96, eliminate DNA contamination with similar efficiency.  However, using the gDNA Eliminator 96 plate of the RNeasy Plus 96 Kit is faster and more economical. 

 

 

FAQ ID -1994
What is the composition of Buffer RLT plus?
In addition to all the components included in Buffer RLT, Buffer RLT Plus — a component of RNeasy Plus Kits — also contains a proprietary blend of detergents. These detergents support the efficient binding of DNA molecules to the gDNA Eliminator column or the gDNA Eliminator plate. The exact composition of Buffer RLT Plus is confidential. Buffer RLT Plus can be purchased separately (cat. no. 1053393).
FAQ ID -2794
What is Tissue-Tek O.C.T., and what is it used for?

Tissue-Tek O.C.T. is an embedding compound for cryosectioning, which is soluble in water. It mainly consists of glycols and synthetic resins. Tissue-Tek O.C.T. is used as matrix for cryosectioning of tissues. Using the O.C.T. the tissue samples can be positioned more easily in the microtome and have better qualities during sectioning.

Most cryosections are fixed using non-crosslinking agents. For isolation of RNA from tissue embedded in Tissue-Tek O.C.T. using non-crosslinking agents the RNeasy Plus Micro Kit or the RNeasy Micro Kit (Protocol: Total RNA Isolation from Microdissected Cryosections), or the RNeasy Mini Kit (RNeasy Mini Protocol for the isolation of Total RNA from Animal Tissue) give great results. We strongly recommend removing as much of the embedding compound as possible prior to RNA extraction from the sections. Please see QIAGEN News article, Issue 1 1998, 'Effects of malnutrition on expression of lactase in children' for successful RNA isolation from O.C.T.-embedded tissue using the RNeasy Mini Kit.

In case crosslinking agents (e.g. formaldehyde or glyoxal-containing) were used for fixation of the tissue for cryosectioning the RNeasy FFPE Kit is the perfect choice. The RNeasy FFPE Kit is especially designed for purifying total RNA from formalin-fixed tissue sections. Special lysis and incubation conditions reverse formaldehyde modification of RNA for improved results in downstream application. The crosslinking causes the RNA to break, resulting in overall smaller molecules, which look like a smear when analyzed on a formaldehyde gel.

In case DNA as well as RNA from precious samples is of interest from the identical sections, please have a look at the AllPrep DNA/RNA Kits for unfixed tissue or the AllPrep DNA/RNA FFPE Kit for fixed tissue.

FAQ ID -530
What is the minimum amount of starting material that can be used with the RNeasy Plus Micro and AllPrep DNA/RNA Micro Kit?

In our labs, we were able to purify RNA using the RNeasy Plus Micro Kit, or DNA/RNA using the AllPrep DNA/RNA Micro Kit at single cell level. However, generally we do not recommend purification from fewer than 10–20 cells, due to stochastics related to transcript copy numbers present in a single cell.

For example, if an RNA transcript is present at only 1 copy per cell, recovery from a purification using e.g., 10 cells or less, would be less than 1 transcript copy per microliter, given the recommended elution volume of 14 µl. 

FAQ ID -1750
Have you observed co-amplification of genomic DNA from RNA templates used in the QuantiTect Whole Transcriptome Procedure?

No, we have never observed coamplification of genomic DNA from RNA templates used in the QuantiTect Whole Transcriptome protocol when using RNA purified with RNeasy Kits without on-column DNase digestion.

 

 

FAQ ID -1619
Effects of low A260/A230 ratios in RNA preparations on downstream applications

The efficiency of downstream applications depends strongly on the purity of the RNA sample used.  Pure RNA should yield an A260/A230 ratio of around 2 or slightly above; however, there is no consensus on the acceptable lower limit of this ratio.  Possible candidates that can increase the A230 include “salt”, carbohydrates, peptides, and phenol (or aromatic compounds in general).  In our experience, the increased absorbance at 230 nm in RNA samples is almost always due to contamination with guanidine thiocyanate, present at very high concentrations in the lysis buffer or extraction reagent used in most RNA purification procedures.

Please find an article discussing the effect of low 260/230 ratios in RNA preparations on downstream applications on page 7 of QIAGEN Newsletter March 15, 2010 . In summary, we found that concentrations of guanidine thiocyanate of up to 100 mM in an RNA sample do not compromise the reliability of downstream applications.

 

 

FAQ ID -2248
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
Can miRNA and other RNAs smaller than 200 nucleotides be isolated with the AllPrep DNA/RNA Micro Kit or RNeasy Plus Micro Kit?

Yes, RNAs smaller than 200 nucleotides, including miRNA, can be isolated using the AllPrep DNA/RNA Micro or RNeasy Plus Micro Kit. Please follow the specialized protocol 'Purification of total RNA containing small RNAs from cells' in Appendix D of the AllPrep DNA/RNA Micro or the RNeasy Plus Micro Handbook, respectively.

Please note that this protocol is for use with cultured cells only, and is not compatible with tissues.

 

FAQ ID -1749
How can I ensure complete genomic DNA removal when using the RNase-Free DNase Set?

To ensure efficient gDNA removal when doing an on-column digest using the RNase-Free DNase Set in combination with RNeasy Mini the following factors are crucial:

  • prevent overloading by adjusting the amount of starting material to no more than the maximum amounts recommended in the RNeasy Mini Handbook
  • ensure complete disruption and homogenization of the starting material as instructed in the section 'Disruption and homogenization of starting materials' of the handbook
  • strictly follow the protocol for on-column DNase Digestion in Appendix D of the RNeasy Mini Handbook (you can let wash buffer RW1 incubate on the column for 3-5 minutes before centrifuging to enhance removal of excess gDNA prior to applying the enzyme)

In the rare case that trace amounts of genomic DNA are still detectable in sensitive downstream applications such as e.g., realtime RT-PCR, an in-solution digest using the RNase-Free DNAase set can be performed. Instructions are presented in Appendix C of the RNeasy MinElute Cleanup Handbook.

Alternatively, a second on-column digest can be carried out in future preparations, immediately following the RW1 wash after the first incubation with DNase.

FAQ ID -1087
What is the composition of Buffer RPE?
The exact composition of Buffer RPE is confidential. Buffer RPE is a mild washing buffer, and a proprietary component of RNeasy Kits. Its main function is to remove traces of salts, which are still on the column due to buffers used earlier in the protocol. Ethanol, which is added by the user just before using the kit for the first time, is an important ingredient of Buffer RPE.
FAQ ID -2797
What sample types can be used with the AllPrep DNA/RNA 96 Kit?

The Allprep DNA/RNA 96 kit is designed for cultured human or animal cells, and for easy-to-lyse human or animal tissues. Tissues compatible with RNeasy Mini, RNeasy Plus Mini, and RNeasy Plus 96 are also compatible with the Allprep DNA/RNA 96 kit.

For more information on compatible kits and sample types, see our Selection Guide for RNA purification.

FAQ ID -1996
What is the composition of Buffer RW1?

The exact composition of Buffer RW1 is confidential. Buffer RW1 is a proprietary component of RNeasy Kits. Buffer RW1 contains a guanidine salt, as well as ethanol, and is used as a stringent washing buffer that efficiently removes biomolecules such as carbohydrates, proteins, fatty acids etc., that are non-specifically bound to the silica membrane. At the same time, RNA molecules larger than 200 bases remain bound to the column.

Note: Buffer RW1 should not be used for isolation of small RNAs, for example, microRNAs or fragmented RNA from formalin-fixed tissues, as these smaller fragments will be washed away.  Buffer RWT should be used instead.

FAQ ID -2796
Can I buy the gDNA eliminator columns supplied in your RNeasy Plus kits separately?

We do not sell the gDNA eliminator columns separately.

FAQ ID - 3390