- Improve gene expression studies with consistent results from experiment to experiment
- Easily isolate viral or total RNA from whole blood
- Save precious time – complete the protocol in less than 1 hour
- Walk away – integrate with established robotic platforms
- Validated with human, bovine, porcine, and ovine whole blood
Magnetic beads offer many benefits for isolating RNA from cells and viral RNA from cell-free samples. Ambion’s MagMAX™ technology binds RNA more efficiently than glass fiber filter methods, resulting in higher RNA yields. Furthermore, the RNA yields are more consistent, both from experiment to experiment, and over a broad range of sample sizes. Another important benefit of MagMAX over glass fiber filter-based methods is the elimination of filter clogging due to cellular particulates in samples. Since only a small volume of magnetic beads is needed for high efficiency binding, the bound RNA can be eluted in just 30–50 µl of elution buffer. This makes the MagMAX magnetic bead technology extremely useful for concentrating RNA from large, dilute samples. In addition, the MagMAX-96 Blood RNA Isolation Kit can be used for total nucleic acid isolation by omitting the DNase treatment.
Isolate Viral and Total RNA from Whole Blood Samples
Whole blood is an attractive sample source for assays involving nucleic acids because it is readily available, easily accessible, and rich in genetic information. Many protocols for RNA isolation from blood require sample pre-processing steps that can lead to sample mix-up and/or cross-contamination. The MagMAX™-96 Blood RNA Isolation Kit eliminates pre-processing and is designed for rapid, high throughput isolation of total and viral RNA from mammalian whole blood in 96 well plates. The procedure is fast (< 1 hour), simple, and well suited for automation. The recovered viral RNA or total RNA can be used directly for quantitative reverse transcriptase PCR (qRT-PCR). The MagMAX-96 Blood kit is optimized both for use manually with multi-channel pipettors and for use with robotic liquid handlers (see Figure 1 schematic).
Detailed guidelines for general automation are included with the kit, and downloadable protocols for the use of this kit with specific liquid handling systems are available at our Automation Resource.
For the highest quality RNA, use whole blood samples that have not been frozen. Approximately 300–500 ng of intact RNA can be consistently purified from 50 µl of human whole blood. Bovine, porcine, and ovine whole blood samples can also be used. Yields vary from species to species (see Figure 2). The recovered RNA is devoid of PCR inhibitors such as proteins and heme commonly found in RNA isolated from blood.
Bovine viral diarrhea virus (BVDV) is a significant problem worldwide that results in economic losses in the beef and dairy industries. Blood is an ideal sample source to test for BVDV. Reproducibility and sample cross-contamination were tested by collecting bovine blood from an animal known to be BVDV-infected and from an animal known to be free of BVDV infection. The purified RNA was subsequently used for qRT-PCR targeting BVDV RNA. The BVDV RNA target amplification results correspond perfectly to the BVDV negative and positive sample well distribution. (see Figure 3). BVDV RNA was recovered consistently and reproducibly, with no sample cross-contamination observed. In addition, BVDV target amplification was highly consistent, with an average Ct of 23.08 +/- 0.3.
A 96 well magnetic ring stand is required to use this kit. Ambion offers two different 96 well magnetic ring stands. For more information about both, please see our Magnetic Stands product page.
For additional processing plates and lids, Ambion recommends Phenix #MPU-8117 (Plate) and Phenix #MPE-8019 (Lid).
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