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RNeasy Protect Animal Blood System

For collecting and stabilizing animal blood and purifying RNA and miRNA

Features

  • Reliable stabilization of transcript levels at blood collection
  • Convenient collection and storage of small blood volumes
  • Rapid purification of high-quality total RNA
  • Effective recovery of miRNA
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RNAprotect Animal Blood Tubes (50 x 100 µl)

Cat. No. / ID: 76544

50 tubes for collection, storage, and transport of 100 µl animal blood samples with RNA stabilization; to be used with the RNeasy Protect Animal Blood Kit
¥18,500
TubeKit
RNAprotect Animal Blood Tube
RNeasy Protect Animal Blood Kit
Quantity
50 x 100 µl
50 x 500 µl
The RNeasy Protect Animal Blood System 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 RNeasy Protect Animal Blood System provides a complete solution for preparing high-quality RNA from animal blood. Blood from mouse, rat, and other small animals is conveniently collected in RNAprotect Animal Blood Tubes, which contain a reagent that immediately stabilizes cellular RNA. Total RNA is then purified using the RNeasy Protect Animal Blood Kit. Please note that the RNeasy Protect Animal Blood Kit and the RNAprotect Animal Blood Tubes are ordered separately.

Alternatively, the RNeasy Protect Animal Blood Kit can be used to obtain microRNA (miRNA), either in a total RNA fraction or in a separate small RNA fraction. For purification of total RNA containing miRNA, Buffer RWT (cat. no. 1067933) is also required. For purification of small RNA (containing miRNA) and large RNA in 2 separate fractions, the RNeasy MinElute Cleanup Kit (cat. no. 74204) is also required. The procedures can be fully automated on the QIAcube Connect.


Performance

Blood samples collected in RNAprotect Animal Blood Tubes can be conveniently shipped at ambient temperature or easily stored in a refrigerator. Samples can be stored at 15–25°C for up to 48 hours, at 2–8°C for up to 2 weeks, or at –20°C for at least 3 months with no significant RNA degradation (see figure " Highly intact RNA"). Collection of animal blood in RNAprotect Animal Blood Tubes prevents changes in transcript levels, ensuring reliable gene expression analysis in applications such as real-time RT-PCR and microarray analysis (see figure " Effective transcript stabilization"). The RNeasy Protect Animal Blood Kit delivers reproducible yields of RNA, and requires less than 30 minutes of hands-on time (see figure " Reproducible RNA yields"). Proven RNeasy technology delivers purified RNA that is highly pure, with virtually no genomic DNA contamination.
See figures

Principle

When blood samples are collected, the gene expression profile can change significantly within minutes. Anticoagulants such as EDTA prevent blood clotting, but do not prevent changes in gene expression due to induced transcription or regulated turnover of mRNA. Collection of animal blood in RNAprotect Animal Blood Tubes prevents changes in transcript levels. Following digestion and homogenization of the stabilized blood, RNA is then purified from the samples using well-established RNeasy silica-membrane technology (see figure " RNeasy Protect Animal Blood flowchart").
See figures

Procedure

Blood samples are collected in RNAprotect Animal Blood Tubes. The tubes contain a reagent that lyses blood cells and stabilizes intracellular RNA. The RNAprotect Animal Blood Tubes are centrifuged to pellet the samples, which are then washed with water and resuspended in Buffer RSB. After digestion in Buffer RBT with proteinase K, the samples are homogenized by centrifugation through QIAshredder spin columns. Ethanol is added to the samples, and the samples are then centrifuged through RNeasy MinElute spin columns. The bound total DNA is subjected to DNase digestion and washed with Buffer RW1, followed by Buffer RPE. Pure RNA is eluted in Buffer REB. The purification of RNA with the RNeasy Protect Animal Blood Kit can be fully automated on the QIAcube. A separate protocol is provided for isolation of total RNA including miRNA and requires wash Buffer RWT (ordered separately; cat. no. 1067933). For purification of small RNA (containing miRNA) and large RNA in 2 separate fractions, the RNeasy MinElute Cleanup Kit is also required.

Applications

Total RNA and miRNA purified with the RNeasy Protect Animal Blood System can be used for many downstream applications, including:

Gene expression analysis
RT-PCR
Quantitative real-time RT-PCR
Differential display
cDNA synthesis

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsNorthern, dot, and slot blotting, end-point RT-PCR, quantitative, real-time RT-PCR, array analysis, next-generation sequencing
Elution volume14 µl
Sample amount100 µl or 500 µl animal blood
Main sample typeTissue, cells
Purification of total RNA, miRNA, poly A+ mRNA, DNA or proteinTotal RNA
ProcessingManual (centrifugation)
FormatSpin column
TechnologySilica technology
YieldRat blood (500 μl): 10–30 µg; Mouse blood (100 μl): 4–8 µg

Resources

Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.
Package Insert (1)
For collection and RNA stabilization of animal blood
Quick-Start Protocols (1)

FAQ

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
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
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
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 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