RNeasy Protect Animal Blood System

用于采集并稳定动物血液,纯化RNA或包含miRNA的总RNA

S_1084_8_GEN_Kitrot

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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
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RNAprotect Animal Blood Tube
RNeasy Protect Animal Blood Kit
Quantity
50 x 100 µl
50 x 500 µl
RNeasy Protect Animal Blood System 旨在用于分子生物学应用。该产品不能用于疾病诊断、预防和治疗。

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • 对动物血液进行可靠的基因表达分析
  • 方便采集并保存小体积量血液
  • 快速纯化高品质的总RNA
  • 高效回收miRNA(需单独购买缓冲液)

Product Details

RNeasy Protect Animal Blood System提供完整的方案,用于从动物血液中纯化高品质RNA。可将小鼠、大鼠或其他小动物的血液简单地收集在RNAprotect Animal Blood Tubes中,该收集管中装有的试剂可迅速稳定细胞RNA。然后应用RNeasy Protect Animal Blood Kit纯化总RNA。

Performance

RNAprotect Animal Blood Tubes收集的血液样本可在室温下方便的运输或在冰箱中简单的储存。样本可在15–25°C条件下保存48小时,2–8°C条件下保存两周,或在–20°C条件下保存至少三个月,不会出现明显的RNA降解(参见"Highly intact RNA")。动物血液收集在RNAprotect Animal Blood Tubes中可避免转录水平的变化,确保诸如real-time RT-PCR和生物芯片分析等应用中可靠的基因表达分析(参见"Effective transcript stabilization")。使用RNeasy Protect Animal Blood Kit纯化的RNA产量重复性好,只需不到30分钟的操作时间(参见"Reproducible RNA yields")。经验证的RNeasy技术纯化的RNA纯度高,没有基因组DNA污染。

Principle

采集血样的时候,基因表达谱可在几分钟内发生明显的变化。EDTA等抗凝血剂可避免血液凝固,但不能避免mRNA诱导的转录或调控引起的基因表达变化。将动物血液收集在RNAprotect Animal Blood Tubes中可避免转录水平的变化。对稳定的血液进行消化和匀浆后,使用RNeasy硅胶膜技术从样本中纯化RNA(参见"RNeasy Protect Animal Blood flowchart")。

Procedure

血液样本收集在RNAprotect Animal Blood Tubes中。收集管中的试剂可裂解血液细胞并稳定细胞内的RNA。将RNAprotect Animal Blood Tubes离心使样本沉淀,沉淀水洗后重悬在Buffer RSB中。样本在Buffer RBT中被蛋白酶K消化后,样本离心通过QIAshredder离心柱后实现匀浆。在样本中加入乙醇后,离心通过RNeasy MinElute离心柱。使用DNase消化总DNA,并依次用Buffer RW1和Buffer RPE漂洗。用Buffer REB洗脱获得纯化的RNA。使用RNeasy Protect Animal Blood Kit纯化RNA可在QIAcube全自动核酸纯化仪上实现完全自动化。使用单独的实验方案可纯化包含miRNA的总RNA,并需使用漂洗液Buffer RWT(需单独订购;货号:1067933)。从分开的两部分样本中纯化小RNA(包含miRNA)和大RNA需使用RNeasy MinElute Cleanup Kit。

Applications

运用RNeasy Protect Animal Blood System纯化的总RNA和miRNA适用于多种下游应用,包括:

基因表达分析
RT-PCR
定量real-time RT-PCR
差异显示
cDNA合成

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsPCR, qPCR, real-time PCR
Elution volume45–70 µl
Sample amount25–100 mg
Main sample typeTissue samples
Purification of total RNA, miRNA, poly A+ mRNA, DNA or proteinRNA
ProcessingManual (centrifugation or vacuum)
Format96-well plate
TechnologySilica technology
YieldVaries

Resources

安全数据表 (1)
Download Safety Data Sheets for QIAGEN product components.
试剂盒操作手册 (2)
Package Insert (1)
For collection and RNA stabilization of animal blood
快速启动实验方案 (1)
Certificates of Analysis (1)
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
What are the 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