RNA Extraction from Blood: Choosing the Best System for Your Gene Expression Studies
|Blood is the only human tissue routinely available for RNA expression studies and blood-based biomarker discovery, because it can be collected by relatively non-invasive procedures and is rapidly regenerated. Efficient and robust methods for RNA stabilization and extraction from blood are required for these types of analyses (see sidebar, The Challenges of Analyzing Blood Samples). To address these needs, Applied Biosystems, now including Ambion, has developed several approaches for extracting RNA from blood. Since there is no “one-size-fits-all” solution for blood-based sample preparation, we have developed multiple approaches that allow researchers to choose the system that most closely meets the needs of their specific projects.|
Figure 1. Major Determinants in Choosing a Sample Preparation Method.
† Use Ambion GLOBINclear™-Human Whole Blood Globin Reduction Kit for the protocol
* See www.ambion.com/techlib/misc/ribo_miRNA.pdf for protocol
§ See www.ambion.com/techlib/misc/leuko_iso.pdf for protocol
RiboPure-Blood Kit (up to 0.5 mL)
Tempus™ Blood RNA Tubes (3.0 mL)
LeukoLOCK™ System (3–30 mL)
RNA is extracted from the captured cells by lysing cells on the filter and purifying the RNA with Ambion MagMAX™ magnetic beads (included). The kit contains TURBO DNase™ enzyme for the efficient elimination of contaminating genomic DNA. The LeukoLOCK method typically yields 10–20 µg of highly pure total RNA from 9 mL of blood (yields vary by donor). The alternate protocol used for miRNA recovery is based on lysis in TRI Reagent® solution followed by silicon filter purification.