Isolation and purification of high quality RNA from cultured cells in tube and 96-well formats.
Efficient removal of contaminating genomic DNA.
No centrifugation, filtration or precipitation required.
Scalable for automation platforms
Northern Blot analysis
Manual and Automation friendly
High RNA Yield
Total RNA yield obtained with RNAPrep was compared to two competitor products. RNAPrep routinely produced 1.5-3.0 times higher yields and demonstrated consistent total RNA recovery over a 5-log range (Figures 1A and 1B).
Figure 1A. Total RNA from 1x105 293T cells was isolated with RNAPrep and two other competitor methods, Q & P. Quantitation was performed by RiboGreen assay. RNAPrep yielded 1.5-3.0 times greater yield than competitors Q & P.
Figure 1B. Total RNA isolated from 100 to 1x106 3638C lymphocyte cells was used in a TaqMan RT-PCR assay for the beta-ac tin transcript. Recovery of total RNA using RNAPrep was linear over a 5 log range.
Quality Nucleic Acid Purification
RNAPrep was compared against two competitor kits for isolated total RNA quality. Analysis of samples by the Agilent 2100 bioanalyzer showed RNAPrep isolated RNA had consistently higher RIN scores (Figure 2) that asses 18/28S ribosomal peak highs and ratios along with several other parameters to provide an objective measure of RNA quality on a scale of 1 to 10 (for more information on RIN RNA quality scores refer to Genomics and Proteomics v.4, no.5, pgs. 14-21).
Figure 2. Total RNA was isolated from 1x105 293T cells using (A) RNA Prep, (B) competitor Qs vacuum filtration isolation kit, or (C) competitor Ps magnetic bead total RNA isolation kit. All were analyzed using the Agilent 2100 bioanalyzer. (D) An equal percentage of eluted RNA from each kit was analyzed by agarose gel electrophoresis using a 1.2% E-gel (Invitrogen). (E) Average RNA Integrity Number (RIN) as determined from Agilent 2100 bioanalyzer, reported on a scale of 1-10, 10 being highest quality.
RNAPrep can be performed either in single tube or in full automation plate formats, making it completely scalable. The flexibility of SPRI magnetic bead technology allows you to run partial plates without incurring costs from running fewer than 96-wells. RNA quality obtained with an automated protocol
is indicated in Figure 3.
Quality Downstream Performance
RNA isolated with RNAprep is suitable for downstream applications such as RT-PCR and microarray gene expression analys is. In Figure 4, RNA isolated with RNAPrep from 1x105 293T cells was used as a template for RT-PCR of human genes of varying abundance levels.
RNAPrep is a simple, 3-step total RNA extraction and purification method that delivers superior performance in yielding high-quality RNA for use in applications such as microarrays and RT-PCR gene expression analysis. The kit provides the benefits of SPRI magnetic bead technology that enable efficient removal of unwanted contaminants without the need for filtration or centrifugation. RNAPreps flexibility and scalability allow you to work in single tube or fully automated plate formats.
Available For Use In Your Laboratory
To Place an Order or Receive Technical Assistance:In the U.S. and Canada call us at 800-361-7780.Outside the U.S. and Canada call us at:+1-978-867-2603.Visit us at www.agencourt.com
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