Arcturus® PicoPure® Frozen RNA Isolation Kit

• Greater sensitivity—isolate RNA from even a single cell
• Superior efficiency—maximize RNA recovery from samples
• Streamlined concentration—elute with low volumes
• Flexible applications—compatible with many tube sizes
• Integrated contamination prevention—column lids help ensure that samples remain RNase-free

The PicoPure® RNA Isolation Kit is engineered to recover high-quality total RNA from fewer than 10 cells. The high recovery rate is important not only for small samples containing picograms of RNA, but also for samples containing up to 100 µg of RNA. Since the RNA elutes in as little as 10 µL of buffer, it is ready for gene expression analysis without any vacuum concentration. This kit is available in two sizes: enough for 40 or 200 isolations (Figure 1).

Figure 1. Isolate RNA from a single cell. Quantitative real-time PCR (qRT-PCR) of RNA isolated from 1 to 1,000 laser capture microdissected (LCM) cells. Cultured mouse Leydig (TM3) cells (ATCC) were spun down onto a microscope slide, fixed, and dehydrated. Replicate samples of 1, 10, 100, and 1,000 cells were microdissected using the PixCell® IIe LCM Instrument. RNA was isolated using the PicoPure® RNA Isolation Kit, reverse-transcribed, and subjected to a qRT-PCR assay for GAPDH. Quantifiable message can be detected from a single cell, and the fluorescence signal is proportional to the cell number for all samples studied.

Small, valuable samples often cannot be replaced. Performing gene expression studies on these precious samples requires getting maximum RNA recovery. Comparative studies show that, with small, RNA-poor cell or tissue samples, the PicoPure® kit dramatically outperforms other commercial total RNA isolation kits in efficiency.

The simple isolation protocol consists of extracting cellular RNA, then loading the extract onto the specially designed MiraCol™ Purification Column to bind the RNA. After washing away impurities, the RNA elutes in only 10 µL of buffer, ready for use without vacuum concentration or transfer into a new reaction tube.

The PicoPure® RNA Isolation Kit features unique, high-recovery MiraCol™ Purification Columns, engineered to maximize recovery from microscopic samples. The columns have convenient integrated lids to help ensure that samples remain RNase-free. In addition, the kit includes all necessary buffers, an easy-to-follow user guide, optimized protocols for handling laser capture microdissection (LCM) samples, and protocols for isolating RNA from larger samples (Figure 2).

Figure 2. MiraCol™ Purification Columns

RNA isolated with the PicoPure® RNA Kit is ready, without vacuum concentration, for amplification with the RiboAmp® RNA Amplification Kit. The two kits together enable highly reproducible gene expression results.

The PicoPure® RNA Isolation Kit preserves mRNA in all abundance classes through the isolation process, minimizing the loss of important genes of interest. Low, medium, and high-abundance genes are recovered and available for expression analysis (Figure 3).

The PicoPure® RNA isolation process allows for high-quality RNA. Total cellular RNA prepared from LCM samples remains intact after isolation, increasing confidence in subsequent molecular analysis (Figure 4).

		Figure 3. RT-PCR of low, medium, and high-abundance genes in RNA isolated using the PicoPure® Kit. RT-PCR was performed on RNA from 500 cells captured from mouse liver and kidney. Equal quantities of cDNA were analyzed with three mouse primer sets: Elongation Factor 1-a (EF-1a, high-abundance gene, ~3,000 copies/cell, 187 bp), Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH, medium-abundance gene, 300–3,000 copies/cell, 357 bp), and Protein Phosphatase 1 (PP1, low-abundance gene, <300 copies/cell, 498 bp). Samples were run on an acrylamide gel. M: molecular weight markers. Lane 1: kidney EF-1a. Lane 2: liver EF-1a. Lane 3: kidney GAPDH. Lane 4: liver GAPDH. Lane 5: kidney PP1. Lane 6: liver PP1. Lane 7: liver EF-1a no RT control. Lane 8: no RT template control.
click to enlarge		Figure 4. The PicoPure® Kit maintains RNA integrity. Total RNA from 15,000 mouse kidney cells was isolated from two samples with the PicoPure® RNA Isolation Kit.RNA samples were run using an Agilent Bioanalyzer 2100. Lane 1: Total RNA from non-LCM cells. Lane 2: Total RNA from LCM-captured cells. Comparable 18S and 28S ribosomal RNA band intensities are observed between RNA isolated from both the intact tissue and the LCM sample, demonstrating high quality of RNA obtained.

The Arcturus® PicoPure® RNA Isolation Kit is for research use only, and is not intended for animal or human therapeutic or diagnostic use.