Biosynthesis has developed a system that enables the collection, stabilization, storage, and transportation of whole blood specimens, together with a rapid and efficient protocol for purification of intracellular RNA.
The system requires the use of Biosynthesis’s Blood RNA Preservative Tubes for blood collection and RNA stabilization, followed by RNA purification using spin column chromatography based on Biosynthesis’s proprietary resin.
Collection of whole blood is the first step in many molecular assays used to study cellular RNA. However, a major problem in such experiments is the instability of the cellular RNA profile in vitro.
Studies have shown that the copy numbers of individual mRNA species in whole blood can change more than 1000-fold during storage or transport at room temperature, due to rapid RNA degradation and induced expression of certain genes after the blood is drawn.
Such changes in the RNA expression profile make reliable studies of gene expression impossible. A method that preserves the RNA expression profile is therefore essential for accurate analysis of gene expression in human whole blood.
Sample Collection and Stabilization
Biosynthesis’s Blood RNA Preservative Tubes (included in kit) feature an aqueous preservative that protects RNA molecules from degradation by RNases and minimizes induction of gene expression. Biosynthesis’s Blood RNA Preservative Tubes are intended for the collection of whole blood and stabilization of cellular RNA for up to 12 days at 18–25 °C or up to 14 days at 2–8 °C.
Studies evaluating stability for longer time periods at other temperatures are ongoing, and further information regarding these findings can be obtained by contacting Biosynthesis Biotek. The actual duration of RNA stabilization may vary depending upon the species of cellular RNA and the downstream application used. Five different transcripts were validated for stabilization specifications (IL-16, NFKβIA, RAS1, FOS and IL1B gene transcripts).
RNA Concentration and Purification
Biosynthesis’s Blood RNA Preservation and Purification Kit provides a rapid method for the isolation and purification of total RNA from 3 mL of human whole blood collected in Biosynthesis’s Blood RNA Preservative Tubes. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA).
The RNA is preferentially purified from other cellular components such as proteins, without the use of phenol or chloroform. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.
Features and Benefits
- Efficient Whole Blood RNA Preservation - Preserve whole blood RNA for up to 12 days at 18–25 °C or up to 14 days at 2–8 °C
- Fast and easy processing - Rapid spin-column format allows for the processing of multiple samples in 30 minutes.
- Isolate a diversity of RNA species - All RNA species can be isolated, from large mRNA and ribosomal RNA down to microRNA (miRNA)
- No phenol:chloroform extractions - Total Preserved blood RNA is isolated without the use of harmful chemicals such as phenol or chloroform
- High Quantity and Quality - No contamination or degradation of preserved blood RNA is observed.
- Quantitative, real-time RT-PCR for both large mRNA and small RNA including miRNA
- RT-PCR for both large mRNA and small RNA including miRNA
- Expression profiling
- Next Generation Sequencing for RNA and miRNA
- microRNA cloning and amplification
- PCR-based virus detection
- Northern blotting
- RNase protection
- Primer extension