PicoPure® RNA Isolation Kit

  • Isolate RNA from Even A Single Cell
  • Get Superior RNA Recovery
  • Eliminate Time-Consuming RNA Concentration
  • Use High-yield Purification Columns
  • Deliver RNA Ready for Amplification and Microarray Analysis
  • Retain Low-abundance mRNA
  • Maintain High RNA Quality



Isolate RNA from Even a Single Cell

The PicoPure® RNA Isolation Kit is engineered to recover high-quality total RNA consistently from fewer than ten cells. The high recovery rate attained using the Kit is valuable 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. The Kit is available in two sizes, enough for 40 or for 200 isolations.



Quantitative Real-Time PCR (QRT-PCR) of RNA isolated from 1 to 1000 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 1000 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 using a LightCycler® (Roche). Quantifiable message can be detected from a single cell, and the fluorescence signal is proportional to the cell number for all samples studied.

Get Superior RNA Recovery

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.



RNA recovery efficiency comparison among commercial RNA isolation kits. Using the PicoPure Kit and other leading products for purifying small RNA samples, 100 ng of Cy3-labeled RNA was bound to columns, washed, and eluted following the corresponding manufacturer's protocol. Values represent average percent recovery of RNA relative to the PicoPure Kit, of five replicates per kit, quantified using fluorescence detection.

Eliminate Time-consuming RNA Concentration

The simple isolation protocol consists of extracting cellular RNA, then loading the extract onto the 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.

Use High-yield Purification Columns

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 comes complete with all necessary buffers, an easy-to-follow user guide, optimized protocols for handling Laser Capture Microdissected (LCM) samples, and protocols for isolating RNA from larger samples.

Deliver RNA Ready for Amplification and Microarray Analysis

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

        

Reproducible amplification and microarray results using PicoPure-isolated RNA. Panel A. Total RNA was isolated from six independent LCM samples using the PicoPure RNA Isolation Kit. RNA from each capture was amplified two rounds using the RiboAmp RNA Amplification Kit. Amplified aRNA was run on an agarose gel. M: marker. Lane 1: positive amplification control. Lane 2: negative control. Lane 3: kidney duct 1. Lane 4: kidney duct 2. Lane 5: mouse kidney glomerulus. Lane 6: whole kidney. Lane 7: SKBR3 cell line (250 captured cells). Lane 8: SKBR3 cell line (500 captured cells). Panels B-C. RNA from 1500 mouse kidney duct cells was isolated using the PicoPure RNA Isolation Kit, amplified two rounds using the RiboAmp Kit, labeled with Cy5-dUTP, and hybridized onto 9000- element mouse cDNA arrays. One region of two arrays illustrates high correlation between gene expression patterns (R = 0.979).

Retain Low-abundance mRNA

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 consistently recovered and available for expression analysis.



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, ~3000 copies/cell, 187 bp), Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH, medium- abundance gene, 300-3000 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.

Maintain Hight RNA Quality

The PicoPure RNA isolation process ensures high-quality RNA. Total cellular RNA prepared from LCM samples remains intact after isolation, increasing confidence in subsequent molecular analysis.

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.

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