Sample Collection & StabilizationDNA5RNA19Tissue/FFPE5RNeasy Protect Bacteria Mini KitPrint For stabilization and purification of up to 100 µg total RNA from bacteriaImmediate in vivo RNA stabilization and protectionReliable gene expression and gene-profiling dataNo need for liquid nitrogen, dry ice, or phenolReady-to-use, high-quality total RNA in minutesNo CsCl gradients, no LiCl or ethanol precipitationThe RNeasy Protect Bacteria Mini Kit includes RNAprotect Bacteria Reagent for stabilizing RNA in bacterial samples, and RNeasy spin columns for purifying up to 100 µg of high-quality RNA using silica-membrane technology. Efficient disruption of bacterial samples can be achieved using the TissueLyserII. The kit can be automated on theQIAcube Connect.Buy ProductsProduct DetailsSpecificationsProduct ResourcesBuy ProductsCat No./ID:74524RNeasy Protect Bacteria Mini Kit (50)$528.00Order ProductRNeasy Mini Kit (50) and RNAprotect Bacteria Reagent (2 x 100 ml)The RNeasy Protect Bacteria Mini Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.Product Details RNeasy Mini spin column.The RNeasy Mini spin column contained in the RNeasy Protect Bacteria Mini Kit.RNAprotect Bacteria Reagent prevents mRNA degradation.In order to monitor mRNA degradation only, transcription was stopped by adding the RNA polymerase inhibitor rifampicin to a growing culture of E. coli. The culture was split into halves, and RNAprotect Bacteria Reagent was added to one half. Samples were left at room temperature for 0, 4, 8, and 16 minutes before centrifugation and RNA isolation. The resulting RNA was analyzed by agarose gel electrophoresis (top panel). The expression of two marker genes with different half-lives was examined by northern blot analysis. Middle panel: ompA (half life of 15 minutes); bottom panel: α-lactamase (half life of 2–5 minutes).RNAprotect Bacteria Reagent procedure.Accurate gene expression profiles.Total RNA was isolated from RNAprotect stabilized E. coli cultures using the RNeasy Protect Bacteria Kit (RNeasy Protect Bacteria) or from unstabilized cultures using a commercial precipitation method (Supplier EIII). To distinguish between gene expression under defined culture conditions and effects of artifactual gene induction during harvesting and RNA isolation, the RNA polymerase inhibitor rifampicin was added to half of the culture prior to RNA isolation. Differences in transcript levels with and without rifampicin therefore generally reflect the degree of RNA degradation. [A] GeneChip analysis of E.coli microarrays was carried out according to standard Affymetrix protocols. [B] The percentage of genes expressed was estimated as the number of different transcripts determined present by GeneChip analysis divided by the total number of transcripts represented on the microarray. (Data from a collaborative study with Affymetrix.)PerformanceDuring traditional methods for cell harvesting and RNA isolation, enzymatic degradation of RNA leads to reduction or loss of many transcripts. The reduction is particularly significant in bacterial mRNA molecules, which have very short half lives of only a few minutes. In addition, genes can be induced during handling and processing of bacterial samples, leading to higher expression. Use of RNAprotect Bacteria Reagent overcomes these problems by providing immediate stabilization prior to RNA isolation (see figures "RNAprotect Bacteria Reagent prevents mRNA degradation" and "Accurate gene expression profiles"). Yield from E. coli (6 x 108 cells) is 100 µg of RNA and the RNA yield from Bacillus subtilis (1 x 108 cells) is 15 µg. RNA purified with the kit is high-quality with A260/A280 ratios of 1.9–2.1 (measured in 10 mM Tris·Cl, pH 7.5).PrincipleThe RNeasy Protect Bacteria Mini Kit provides a complete RNA protection and isolation solution, from sample harvesting to pure RNA, in one kit. Proven RNeasy technology, combined with the RNA-stabilizing properties of RNAprotect Bacteria Reagent, allows purification of high-quality, intact RNA. This process efficiently preserves the expression profile of the bacteria at the time of harvesting to ensure reliable gene expression analysis. Following stabilization, RNeasy technology simplifies total RNA isolation by combining the stringency of guanidine-isothiocyanate lysis with the speed and purity of silica-membrane purification (see figure "RNeasy Mini spin column").Procedure Two volumes of RNAprotect Bacteria Reagent are added directly to 1 volume of bacterial culture (≤7.5 x 108 bacteria) prior to RNA isolation, providing immediate stabilization of RNA (see flowchart "RNAprotect Bacteria Reagent procedure"). The stabilization allows time for efficient bacterial lysis using a choice of protocols: enzymatic lysis, mechanical disruption, or a combination of both methods. We recommend the TissueLyser II for efficient mechanical disruption. Ethanol is then added to the lysate to provide ideal binding conditions. The lysate is loaded onto the RNeasy silica membrane (binding capacity 100 µg RNA). Following RNA binding, all contaminants are efficiently washed away. Pure, concentrated RNA is eluted in 30–100 µl of water. The RNeasy Protect Bacteria Mini Kit can be automated on the QIAcube. Amounts of RNA isolated from bacteria can vary due to species and growth conditions. The RNeasy Protect Bacteria Mini Kit is suitable for use with a wide range of bacterial species, both Gram positive (e.g., Staphylococcus aureus and Mycobacterium avium) and Gram negative (e.g., Escherichia coli and Salmonella typhimurium). Bacteria grown in either minimal or complex medium can be used. Since the RNeasy procedure enriches for RNA species >200 nucleotides, RNA yield does not include 5S rRNA, tRNAs, or other low-molecular-weight RNAs. RNeasy Protect Bacteria Kits provide the highest-quality RNA with minimum copurification of DNA. For certain RNA applications that are sensitive to very small amounts of DNA, the residual amounts of DNA remaining can be removed using the RNase-Free DNase Set for convenient on-column DNase treatment during the RNeasy procedure. Applications RNA purified with the RNeasy Protect Bacteria Mini Kit is ideal for use in all applications. Downstream applications include: Northern, dot, and slot blotting End-point RT-PCR Quantitative, real-time RT-PCR Array analysis SpecificationsFeaturesSpecifications ApplicationsPCR, qPCR, real-time RT-PCR, microarray Elution volume30–100 µl FormatSpin column Main sample typeBacteria ProcessingManual (centrifugation) Purification of total RNA, miRNA, poly A+ mRNA, DNA or proteinRNA Sample amount15 –100 µg StabilizationYes TechnologySilica technology Time per run or per prep20 minutes Yield8–70 µgProduct ResourcesYou are not authorized to download the resourceBrochures & Guides (3) Sort optionsSort alphabetically All insights start with the sample: Your comprehensive guide for isolating top-quality RNAShow details(EN) - Stabilization Solutions for Reliable Gene Expression AnalysisShow details(EN) - Analyzing Gene Expression and RegulationShow detailsShow allShow lessFAQs (34) Sort optionsSort alphabetically How do I safely inactivate biohazardous flow-through material?FAQ ID -12ViewHow should I quantify RNA isolated with RNeasy Kits?FAQ ID -32ViewWhy does my isolated RNA have a low OD 260/280 ratio?FAQ ID -97ViewHow can I avoid little or no RNA yields when using an RNeasy Kit?FAQ ID -28ViewWhy do I have to add beta-mercaptoethanol (beta-ME) to lysis Buffer RLT of the RNeasy Kits?FAQ ID -101ViewHow should RNeasy Kits be stored and how long are they stable?FAQ ID -103ViewHow do you ensure that RNeasy buffers are RNase-free?FAQ ID -113ViewWhat is the difference between disruption and homogenization in the RNeasy System?FAQ ID -139ViewAre RNeasy spin columns sold separately?FAQ ID -159ViewHow can I check for purity of RNA isolated using RNeasy Kits?FAQ ID -1023ViewHow can I check the integrity of RNA purified using RNeasy Kits?FAQ ID -1024ViewWhich kit should I use for RNA isolation from Cartilage?FAQ ID -1026ViewDo I have to discard Buffer RLT with beta-Mercaptoethanol (ß-ME) added to it after 1 month of storage?FAQ ID -1037ViewWhat is the difference between Buffers RLT and RLT Plus?FAQ ID -1043ViewHow can I ensure complete genomic DNA removal when using the RNase-Free DNase Set?FAQ ID -1087ViewWhat is the maximum binding capacity of RNeasy spin columns?FAQ ID -290ViewEffects of low A260/A230 ratios in RNA preparations on downstream applicationsFAQ ID -2248ViewWhat is the composition of Buffer RLT?FAQ ID -2793ViewWhat is the composition of Buffer RW1? FAQ ID -2796ViewWhat is the composition of Buffer RPE? FAQ ID -2797ViewHow much RNA does a bacterial cell contain? FAQ ID -2949ViewWhat happens if I spin my lysate on the RNeasy Spin Columns at maximum speed?FAQ ID -514ViewWhat has to be done to an RNA sample before loading it onto an Agilent Bioanalyzer?FAQ ID -528ViewDoes DEPC harm RNA?FAQ ID -529ViewIs it possible to isolate both RNA and recombinant 6xHis-tagged protein from the same sample?FAQ ID -532ViewHow long can I store bacterial culture stabilized in RNAprotect Bacteria Reagent?FAQ ID -614ViewCan I stabilize RNA in bacteria grown on solid media or substrate using the RNAprotect Bacteria Reagent?FAQ ID -615ViewCan the RNase-Free DNase Set be used for DNase digestions of RNA in solution?FAQ ID -619ViewDo you have a kit for RNA isolation from any kind of sample type?FAQ ID -627ViewWhat can be used as an alternative to the A260 measurement for quantification of small amounts of RNA and DNA?FAQ ID -728ViewI accidentally stored Buffer RDD of the RNase-Free DNase Set at°C. Will it still function? -20ViewI ran my RNA out on an agarose gel and can see lots of bands similar to a ladder. Why? FAQ ID -745ViewIs RNAprotect Bacteria Reagent compatible with the RNeasy Mini Kit?FAQ ID -797ViewDo I need to use RNase inhibitors with the RNeasy Kits?FAQ ID -813ViewShow allShow lessQuick-Start Protocols (1) Sort optionsSort alphabetically RNAprotect Bacteria Reagent and RNeasy Protect Bacteria Kits (EN)Show detailsShow allShow lessSafety Data Sheets (1) Sort optionsSort alphabetically MSDS RNeasy Protect Bacteria Mini Kit (50)Show allShow less References 0 fragment fix placeholderCustomers who bought these products also boughtCat No./ID:339452miRCURY LNA miRNA ISH Optimization Kit (FFPE) 2miRCURY LNA miRNA ISH Optimization Kit 2 (miR-21); includes controls, buffer and proteinase K for 400 slides; detects hsa-miR-21-5p$2,231.00Add To CartCat No./ID:1022064GFP-22 siRNA (5 nmol)Control siRNA targeting green fluorescent protein$182.00Add To CartCat No./ID:27500-4-EP-BPPowerBead Plates, Glass (4)MagAttract PowerMicrobiome DNA/RNA EP Kit Components$311.00Add To CartCat No./ID:204574Rotor-Gene Probe RT-PCR Kit (400)For 400 x 25 µl reactions: 3 x 1.7 ml 2x Rotor-Gene Probe RT-PCR Master Mix, 100 µl Rotor-Gene RT Mix, 2 x 2 ml RNase-Free Water$689.00Add To CartCat No./ID:76506RNAprotect Bacteria ReagentRNAprotect Bacteria Reagent (2 x 100 ml)$283.00Add To CartCat No./ID:79254RNase-Free DNase Set (50)1500 Kunitz units RNase-free DNase I, RNase-free Buffer RDD, and RNase-free water for 50 RNA minipreps$113.00Add To CartCat No./ID:14900-50-NF-BTPowerWater DNA Bead Tube (50)Bead Tubes for the DNeasy PowerWater Kit$246.00Add To CartCat No./ID:19131QIAGEN Proteinase K (2 ml)2 ml (>600 mAU/ml, solution)$102.00Add To CartCat No./ID:205311QuantiTect Rev. 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