FFPE microRNA/DNA Rapid Purification Kits,FFPE microRNA/DNA Rapid Purification Kit

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FFPE microRNA/DNA Purification Kits Offered in two sizes (50 preps & 100preps),RP5311 & RP5312 Sequential isolation and purification of RNA and genomic DNA from FFPE tissue samples Advantages Fast and easy processing using rapid spin-column format High yields and quality of nucleic acids Separate fractionation of RNA and DNA Isolate total RNA, from large rRNA down to microRNA (miRNA) No phenol or chloroform extractions FFPE RNA/DNA Purification Plus Kit provides a rapid method for the sequential isolation and purification of total RNA (including microRNA) and genomic DNA from formalin-fixed paraffin-embedded (FFPE) tissue samples. Using formalin to fix tissues leads to crosslinking of the nucleic acids and proteins, and the process of embedding the tissue samples can also lead to fragmentation of the nucleic acids over time. Bioteke's FFPE RNA/DNA Purification Plus Kit provides conditions that allow for the partial reversing of the formalin modifications, resulting in a high quality and yield of nucleic acids. The kit is able to purify all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA), depending on the age of the FFPE tissue as the degree of fragmentation of the RNA will increase over time. The RNA is purified from other cellular components without the use of phenol or chloroform. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays. The purified genomic DNA is also of the highest quality, and can be used in PCR reactions, sequencing, Southern blotting and SNP analysis. Bioteke's Purification Technology Purification is based on spin column chromatography usingproprietary resin as the separation matrix. The nucleic acids are preferentially purified from other cellular components without the use of phenol or chloroform. The process first involves deparaffinization of the FFPE samples through a series of xylene and ethanol washes. Next, the FFPE samples are digested with the provided Proteinase K and Digestion Buffer A using an incubation time which is specific for the recovery of RNA (please see the flow chart on page 3). The soluble lysate containing the RNA is then collected for RNA purification while the remaining sample is further digested for DNA. Buffer RL and ethanol are then added to the lysate containing RNA or DNA, and the solution is loaded onto an RNA Purification Micro Column or a DNA Purification Micro Column, respectively. Bioteke's resin binds nucleic acids in a manner that depends on ionic concentrations, thus only the RNA or DNA will bind to the column while the contaminants will be removed in the flowthrough or retained on the top of the resin. The bound nucleic acid is then washed with the provided Wash Solution in order to remove any impurities, and the purified nucleic acid is eluted with the Elution Solution A or Elution Buffer F. Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. The DNAse I and Proteinase K should be stored at -20C upon arrival. These reagents should remain stable for at least 1 year in their unopened containers. Precautions and Disclaimers This kit is designed for research purposes only. It is not intended for human or diagnostic use. Ensure that a suitable lab coat, disposable gloves and protective goggles are worn when working with chemicals. For more information, please consult the appropriate Material Safety Data Sheets (MSDSs). These are available as convenient PDF files online at www.norgenbiotek.com. The Buffer RL contains guanidinium salts, and should be handled with care. Guanidinium salts form highly reactive compounds when combined with bleach, thus care must be taken to properly dispose of any of these solutions Customer-Supplied Reagents and Equipment You must have the following in order to use the FFPE RNA/DNA Purification Plus Kit: # Benchtop microcentrifuge # 50 - 55oC Incubator or Water Bath # 80Incubator or Water Bath # 90Incubator or Water Bath # 96 - 100% ethanol # Xylene, histological grade # Ice Bucket # # Working with RNA RNases are very stable and robust enzymes that degrade RNA. Autoclaving solutions and glassware is not always sufficient to actively remove these enzymes. The first step when preparing to work with RNA is to create an RNase-free environment. The following precautions are recommended as your best defense against these enzymes. # The RNA area should be located away from microbiological work stations # Clean, disposable gloves should be worn at all times when handling reagents, samples, pipettes, disposable tubes, etc. It is recommended that gloves are changed frequently to avoid contamination # There should be designated solutions, tips, tubes, lab coats, pipettes, etc. for RNA only # All RNA solutions should be prepared using at least 0.05% DEPC-treated autoclaved water or molecular biology grade nuclease-free water # Clean all surfaces with commercially available RNase decontamination solutions # When working with purified RNA samples, ensure that they remain on ice during downstream applications

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BioTeke Corporation

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