Ambion's RiboPure™-Blood procedure overcomes the challenges involved in isolating intact RNA from whole blood.
Dawn Obermoeller, Ambion, Inc.
- From blood to RNA in less than an hour--no need to isolate white blood cells
- RNAlater included to stabilize RNA
- Samples can be stored for days at ambient temperature
- Recovers highly pure RNA, suitable for demanding applications
Ambion's RiboPure™-Blood procedure overcomes the challenges involved in isolating intact RNA from whole blood. In the past, researchers typically had to isolate white blood cells (WBC) from whole blood prior to RNA isolation to deal with the plethora of contaminants present in blood. One of the challenges involved in isolating RNA from blood is that white blood cells make up only a small fraction (about 0.2%) of blood cells. Red blood cells, which account for the vast majority of cells present in blood, contain large amounts of hemoglobin, which inhibits RT-PCR. Plasma, which comprises ~60% of blood, has a high concentration of RNases, as do some white blood cell subtypes, making it necessary to quickly inactivate RNases in blood samples to preserve the RNA. The large amount of hemoglobin in whole blood further complicates RNA purification by clogging filters, beads and other solid-phase matrices, and if carried over will interfere with downstream applications. The RiboPure-Blood Kit combines two sequential RNA purification techniques (Figure 1) to remove all of these potential contaminants, resulting in high yields of pure RNA directly from whole blood (without prior fractionation of WBCs) that is ideal for RT-PCR (Figure 2), microarrays, or any other application with stringent purity needs.
Figure 1. Schematic of RiboPure™-Blood Procedure.
Figure 2. Real-time RT-PCR from RiboPure™-Blood Isolated RNA. Real-time RT-PCR experiment in which p53, BTK, and GAPDH transcripts were detected in the RNA isolated using the RiboPure-Blood procedure. No genomic DNA was detectable in "no RT" control reactions.