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Download HUMAN PRIMARY CELLS RNA PRODUCTS Total RNA
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Page 1 of 2 includes a Certificate of Analysis, which shows the RNA purity using the 260nm/280nm absorbance ratio along with an electropherogram and converted gel image that shows the intact RNA species, such as 18s and 28s rRNA. 5858 Horton Street, Suite 360 Emeryville, CA 94608 Phone: 510-450-3900 Fax: 510-450-3903 www.allcells.com Total RNA Sample HUMAN PRIMARY CELLS RNA PRODUCTS Index Total RNA Comprehensive RNA Shipping and Storage AllCells human primary cell total RNA and comprehensive RNA are shipped in a 1.5ml microtube and shipped on dry ice. RNA is stable at 80°C for more than 6 months, and AllCells recommends an expiration date for RNA one year from delivery when stored properly in DEPCtreated water. Repeated freezing or thawing should be avoided. RNA can be stored at -20°C for a short period of time, but should not be kept in a defrost freezer. This is an Agilent 2100 Bioanalyzer electropherogram image of a sample of Total RNA. The above image shows intact 28s and 18s rRNA signals. Frequently Asked Questions Q. Is there genomic DNA contamination in the RNA products? CAUTION The products purchased from AllCells, LLC are “For In Vitro Research Use Only.” In compliance with U.S. Food and Drug Administration (FDA) Regulations, these products are intended neither for use in clinical diagnostic procedures nor in therapeutic procedures. These products must be: 1. Treated as potentially contaminated biological specimens even if available serological reports are negative. 2. Handled by establishing or following appropriate safety control procedures to ensure the safety of using these products. Total RNA Product Description AllCells human primary cell Total RNA is highly purified from viable, human cells. Those cells are isolated from human blood, bone marrow, or umbilical cord blood using Miltenyi’s magnetic automatic cell sorting (MACS) technology. The purity of the cells is analyzed by FACS assay and is higher than 90%. Application AllCells human primary cell Total RNA is developed to meet research needs in gene expression and cloning studies in the Hematology and Immunology field. It can be used in microarray gene expression profile study, cDNA synthesis, RT-PCR, Northern blot, RNA protection assay, dot blot, and other molecular analysis. A. AllCells Total RNA products have been treated with DNase I to minimize genomic DNA presence. However, no procedure can guarantee absolute absence of all genomic DNA traces. Q. Is there micro RNA in the Total RNA products? A. No, there is no micro RNA in the Total RNA products. However, AllCells Comprehensive RNA products contain the small and micro RNA. Q. What should I expect in terms of quality and integrity of the total RNA? A. At AllCells, we analyze the RNA quality and integrity by running the RNA product on an Agilent 2100 Bioanalyzer, which will provide a RIN, or RNA Integrity Number, which should usually be between 9.0 and 10.0 for indication of good integrity. The Agilent electropherogram image also allows us to observe the 18s and 28s ribosomal RNA bands, which are another indicator of good RNA quality. Q. Why does my RNA product come in DEPC-treated water? A. AllCells stores and ships RNA products in DEPC-treated water which is nuclease-free, and can maintain quality and integrity of the RNA for at least 6 months to a year in –80°C. DEPC-treated water does not interfere with any downstream (post-purification) assays. For extended storage, the RNA can be precipitated and re-dissolved in Tris-EDTA (1x). Total RNA Extraction Freshly purified cells are lysed in Rnase-free conditions. Total RNA is isolated with an optimized RNA purification method. Residual genomic DNA is further removed with RNase-free DNase I. RNA Purity The high quality of AllCells total RNA is validated through an Agilent Bioanalyzer 2100 (Agilent Technology, Inc.). In each package, AllCells 5000-FRM-508. Rev. no. 006 January 2009 © 2008 AllCells, LLC. All rights reserved. Page 2 of 2 COMPREHENSIVE RNA Product Description AllCells Comprehensive RNA includes a combination of Total RNA (>200nt) and small and micro RNA (19-200nt). AllCells Total RNA product is co-purified together with small RNA and micro RNA populations. The below image is a sample of an Agilent 2100 Bioanalyzer converted gel image from various cell types. Background Recent discoveries have shown that micro RNA has a significant cellular presence and role as repressors of gene expression. Micro RNA has also been found to be a potential factor in cancer and heart diseases. Comprehensive RNA Extraction AllCells uses an optimized phase extraction method to isolate the freshly * purified cells, which are lysed with TRIZOL® reagent. RNA Purity The Comprehensive RNA is guaranteed to have the highest purity and integrity with every lot of the product validated by an Agilent 2100 Bioanalyzer. Every qualifying lot of product is shown to have an intact tRNA, 5s, 5.8s, 18s, and 28s rRNA and an extremely minimal presence of genomic DNA. Small and Micro RNA Related Products Customers who have ordered AllCells Total or Comprehensive RNA are also interested in: cDNA: a reverse transcriptase copy from Total RNA primed with oligo-d(T). The original RNA templates are removed from cDNA by using RNase H. Lysates: AllCells fresh cells are spun down and lysed with RLT or Trizol. For more information on AllCells, LLC products, please contact an AllCells Customer Service Representative at the following: Phone: 510-450-3900; Toll-Free: 1-888-535-3444; Fax: 510-450-3903 Email: [email protected]; Web: http://www.allcells.com This is a Agilent 2100 Bioanalyzer electropherogram image of a sample of small and micro RNA regions. The image shows intact signals of tRNA (~40-70nt) and 5s (~80-100nt) and 5.8s (~130-140nt) rRNA. The presence of micro RNA (~19-40nt) is evident in the above image. COMPREHENSIVE RNA (large scale) * TRIZOL is a registered trademark of Invitrogen. The electropherogram image above shows the profile of Comprehensive RNA on a larger scale. Intact 18s and 28s rRNA bands are observed alongside a condensed signal from the small and micro RNA regions. 5000-FRM-508. Rev. no. 006 January 2009 © 2008 AllCells, LLC. All rights reserved.