PreAnalytiX — Integrated, Standardized Systems

The joint venture PreAnalytiX combines the expertise of BD and QIAGEN in sample collection and nucleic acid isolation, respectively. Products from PreAnalytiX provide integrated and standardized systems for the collection and stabilization of clinical samples, together with efficient methods for nucleic acid isolation. QIAGEN is the distributor for PreAnalytiX products.

PAXgene Blood RNA System

The PAXgene Blood RNA System is an integrated and standardized system for the collection and stabilization of whole blood specimens and isolation of cellular RNA. The system requires the combined use of PAXgene Blood RNA Tubes for blood collection and RNA stabilization, followed by RNA isolation using the PAXgene Blood RNA Kit. The system is for research use only, and not for use in diagnostic procedures.

Features and benefits

The first integrated system for collection, stabilization, and purification of intracellular RNA from whole blood
Immediate stabilization of intracellular RNA, providing in vivo snapshots of geneexpression profiles
RNA stabilization for up to 5 days at room temperature during blood storage and transport
Standardized sample processing prior to analysis
No organic extraction or precipitation
Isolation of high-quality cellular RNA

Principle

In the PAXgene Blood RNA System, blood is collected directly into PAXgene Blood RNA Tubesand RNA is isolated using the PAXgene Blood RNA Kit. PAXgene Blood RNA Tubes contain a proprietary blend of reagents that bring about immediate stabilization of RNA. This blend prevents the drastic changes in the cellular RNA expression profiles that normally take place in vitro, after blood collection (see figures "Stabilization of Gene Expression Using the PAXgene Blood RNA System" and "Detection of Tyrosinase Following Blood Storage"). Blood samples can be stored in the PAXgene Blood RNA Tubes for up to 5 days at room temperature. RNA is then isolated using silica-gel–membrane technology supplied in the PAXgene Blood RNA Kit. The resulting RNA accurately represents the expression profile in vivo and is suitable for use in a range of downstream applications.

Procedure

The PAXgene Blood RNA procedure is shown in the flowchart. Blood samples (2.5 ml) are collected in PAXgene Blood RNA Tubes, and may be stored or transported at room temperature if desired. RNA isolation begins with a centrifugation step to pellet nucleic acids in the PAXgene Blood RNA Tube. The pellet is washed, and Proteinase K is added to digest proteins. Alcohol is added to adjust binding conditions, and the sample is applied to a PAXgene RNA spin column. During a brief centrifugation, RNA is selectively bound to the PAXgene silica-gel membrane as contaminants pass through. Following washing steps, RNA is eluted in an optimized buffer.

Stabilization of Gene Expression Using the PAXgene Blood RNA System

Blood was collected and RNA was isolated using either A standard methods (collection in EDTA tubes; no stabilization; RNA isolation using a guanidinium-based method), or B the PAXgene Blood RNA System (for RNA stabilization and isolation). The graphs show changes in expression of 12 genes after blood collection, measured using real-time RT-PCR. (Data kindly provided by Source Precision Medicine, Boulder, Colorado, USA.)

Detection of Tyrosinase Following Blood Storage

Blood samples were collected from 6 different donors, spiked with tumor cells expressing tyrosinase, and stored for up to 10 days at room temperature. RT-PCR was used to measure levels of tyrosinase. A 1 cell per 2.5 ml blood; B 10 cells per 2.5 ml blood. WB: tyrosinase detection in RNA isolated from whole blood collected and stored in standard EDTA tubes. PAX: tyrosinase detection using the PAXgene Blood RNA System for sample collection, and RNA stabilization and isolation. (Data kindly provided by LMO, Munich, Germany.)

Downstream applications

Typical yields of RNA isolated from 2.5 ml human whole blood are between 4 and 20 µg. The isolated RNA is ready for use in a wide range of downstream applications, including:

Nucleic acid amplification technologies, such as RT-PCR and NASBA
Expression-array and expression-chip analysis
Quantitative RT-PCR, including TaqMan technology
cDNA synthesis
RNase and S1 nuclease protection
Northern, dot, and slot blot analysis
Primer extension

PAXgene Blood RNA Tubes and Kits

For blood collection, and RNA stabilization and isolation

操作手册:
PAXgene Blood RNA Kit Handbook (PDF version, 217KB)

PAXgene Blood RNA Tube Product Circular (PDF version, 188KB)

Ordering Information
Product	Contents	Cat. No.
PAXgene Blood RNA Tubes (100)	100 Blood Collection Tubes. To be used in conjunction with the PAXgene Blood RNA Kit (50).	762125

PAXgene Blood RNA Validation Kit (10)	10 Blood Collection Tubes, 10 PAXgene Spin Columns, Processing Tubes, RNase-free Reagents and Buffers	762132

PAXgene Blood RNA Kit (50)	50 PAXgene Spin Columns, Processing Tubes, RNase-free Reagents and Buffers. To be used in conjunction with the PAXgene Blood RNA Tubes.	762134

Accessories to be ordered from BD: BD Vacutainer Brand Safety-Lok Blood Collection Set, Cat. No. 367281 (USA only); 367286 (other countries); BD Vacutainer Standard Needle Holder, Cat. No. 364888.