Sample Preparation - J&B Lab. S.A.C
Sample PreparationRNA, DNA, and Protein Sample PreparationRNA Kits and ReagentsDNA Kits and ReagentsSample Cleanup and PurificationProtein ExtractionProtein Sample Cleanup, Reduction,and AlkylationProtein FractionationProtein Sample DepletionSurface-Enhanced Laser Desorption/Ionization225689101112Ordering Information13
RNA, DNA, and Protein Sample PreparationRNA Kits and Reagentswww.bio-rad.com/nasampleprepRNA, DNA, and Protein Sample PreparationRNA Kits and ReagentsBio-Rad offers a broad range of sample preparation products for RNA isolation. Scalable and high throughput productsare available.See AlsoAmplificationreagents:pages 305–310.Real-time PCRdetection systems:pages 300–302.Experion automatedelectrophoresissystem:pages 112–116.RNA Isolation Product Selection GuideMiniAurum Total RNA KitsFatty and Fibrous Tissue96PureZOL RNAIsolation ReagentFormatMini columnFiltration (vacuum or spin)96-well plateFiltration (vacuum or spin)Single solution organicextraction1 x 1068 x 1082 x 107——1 x 1072.4 x 1093 x 107100 mg100 mgMini columnFiltration (vacuum or spin)Maximum startingmaterial amountsCultured cells2 x 1061 x 107Bacterial cells2.4 x 1092.4 x 109Yeast cells3 x 1073 x 107Hard animal tissue 20 mg100 mgSoft to moderately 40 mg100 mghard animal tissuePlant tissue40 mg100 mg—100 mgIsolation methodSilica membraneLysis with PureZOL reagent,purification on silica membraneSilica membraneOrganic extractionNumber of preps5050192 (2 x 96-well plate)50 or 100 (1 ml/prep)Number of washes333—DNase I included*YesYesYesNoDNase I digest time15 min (animal tissue, 25 min)15 min10 min—Total preparation time** 50–80 min (with DNase I digest) 50–80 min (with DNase I digest) 60 min (with DNase I digest) 60 minBinding capacity 100 µg 100 µg 40 µg—Maximum elutionvolume80 µl (2 x 40 µl elutions)2 x 40 µl80 µl30–100 µlYield***20–30 µg/2 x 106 NIH 3T3 cells140–150 µg/1 x 107 293H cells10 µg/1 x 106 NIH 3T3 cells20–40 µg/2 x 106 HeLa cells90–100 µg/100 mg brain17 µg/1 x 106 HeLa cells15–20 µg/40 mg brain50–60 µg/100 mg breast11 µg/2 x 107 S. cerevisiae cells30–55 µg/20 mg thymus80–90 µg/100 mg heart5 µg/8 x 108 E. coli cells15–30 µg/40 mg spleen60–70 µg/100 mg skin5 µg/8 x 108 B. cereus cells8–15 µg/40 mg liver50–60 µg/100 mg cartilage15–20 µg/40 mg kidney20–30 µg/2.4 x 109 E. coli cells8–15 µg/40 mg lung60–65 µg/3 x 107 S. cerevisiae cells15–30 µg/40 mg potato90–100 µg/100 mg potato20–30 µg/3 x 107 S. cerevisiae cells 5–10 µg/100 mg Arabidopsis35–55 µg/2.4 x 109 E. coli cells10–15 µg/50 mg A. niger35–55 µg/2.4 x 109 B. cereus cells1.37 µg/1 x 105 293H cells1.15 µg/mg brain1.37 µg/mg fat1.2 µg/mg heart0.66 µg/mg cartilage1.2 µg/mg skin1.0 µg/mg potato0.1 µg/mg Arabidopsis0.38 µg/mg A. niger0.63 µg/2.4 x 107 E. coli cells0.31 µg/3 x 105S. cerevisiae cellsPurityA260 /A280 of 1.9–2.1A 260 /A 280 of 1.9–2.1A 260 /A280 of 1.9–2.1A 260 /A 280 of 1.9–2.1* Removal not required.** Total preparation time will vary depending on the tissue or cell type and on which format is used (vacuum or spin).*** Yields will vary depending on the developmental stage, tissue type or cell line, and growth conditions used.2 Sample Preparation
RNA, DNA, and Protein Sample PreparationRNA Kits and Reagentswww.bio-rad.com/nasampleprepAurum Total RNA KitsOrder Info: Pg 13PureZOL reagent:page 4.Aurum total RNA kits are designed and formulated toassist in the isolation of highly pure and intact RNA from avariety of starting materials. Isolated RNA is compatible withdownstream applications including real-time qPCR, northernblotting, microarray analysis, and cDNA library construction. See AlsoTransfection reagents:pages 283–284.Nucleic acidamplification:pages 296–319.Isolate high yields of RNA from a wide range of sampletypes in less than 60 minDNase treatment ensures genomic DNA removalProduce ready-to-use RNA suitable for the mostdemanding downstream applicationsAurum Vacuum ManifoldAurum total RNA kits offer streamlined processing using achoice of vacuum- or spin-mediated protocols. If vacuummediated purification is preferred, the protocol may becarried out on the Aurum vacuum manifold.For More InformationWeb: www.bio-rad.com/rna-isolationRequest or download bulletins: 2920, 5282, and 2919Aurum Total RNA Mini KitThe Aurum total RNA mini kit produces DNA-free totalRNA from a wide range of starting materials, includingcultured cells, bacteria, and yeast, as well as animal andplant tissues. The kit utilizes a stringent reagent composedof guanidine isothiocyanate and b-mercaptoethanolfor efficient sample lysis and quick RNase inactivation,followed by purification on silica membrane in a spincolumn format using a spin- or vacuum-mediated protocol.The kit can also be used for RNA cleanup and desalting.Aurum Total RNA Fatty and Fibrous Tissue KitThe Aurum total RNA fatty and fibrous tissue kit producestotal RNA from samples that are difficult to disrupt. Thekit is effective for purifications involving fatty and fibroustissues or samples, such as fungi, that are rich in RNases.The kit utilizes PureZOL RNA reagent, a potent phenolbased reagent, which effectively lyses tissues and cells.The kit includes spin columns that can be processed usinga choice of vacuum- or spin-mediated protocols.Aurum Total RNA 96 KitThe Aurum total RNA 96 kit reproducibly isolates DNA-freetotal RNA from cultured cells, bacteria, and yeast in under60 min in a 96-well format for high-throughput total RNAisolation. The total RNA binding plate is designed for useon the Aurum vacuum manifold.Aurum Total RNA Fatty and Fibrous Tissue KitAurum Vacuum ManifoldVacuum-mediated RNA purifications can be carried outin the Aurum vacuum manifold. The manifold serves as avacuum purification platform for either 96-well plates orindividual spin columns. Benefits include: Processing of up to 18 spin columns at one time niversal manifold that converts from plate to columnUformat by inserting the column adaptor plateConsumables and AccessoriesThe reagents and plasticware used in the Aurum RNAkits are available as refill orders. These products can beordered separately from the Aurum total RNA kits.Sample Preparation 3
RNA, DNA, and Protein Sample PreparationRNA Kits and ReagentsSee AlsoAurum total RNA kits:page 3.www.bio-rad.com/nasampleprepPureZOL RNA Isolation ReagentOrder Info: Pg 13The PureZOL RNA isolation reagent protocol is animprovement over the rapid, widely used, and provenmethod of RNA isolation developed by Chomczynski andSacchi (1987). PureZOL RNA isolation reagent is a potentmonophasic combination of phenol and the chaotropic agentguanidine isothiocyanate, which effectively lyses cells andtissues, deproteinates RNA, and inactivates endogenousnucleases in a single step. DNA and protein are efficientlyremoved from the RNA following phase separation.The ready-to-use PureZOL RNA isolation reagent is a versatileand efficient means of isolating high yields of RNA from avariety of sources including cultured cells, animal and planttissue, yeast, virus, and bacteria samples. The single-solutionformat permits recovery of RNA from small quantities oftissues or cells, making it ideally suited for gene expressionanalysis or whenever sample quantities are limited. TotalRNA isolated using PureZOL RNA isolation reagent is free ofDNA and protein and can be used for northern blot analysis,in vitro translation, poly(A) selection, RNase protectionassays, RT-PCR, and molecular cloning. Since no spincolumns are used in the protocol, the protocol is scalable toaccommodate a wide range of sample sizes.For More InformationWeb: www.bio-rad.com/rna-isolationiScript RT-qPCR Sample Preparation Reagent 104103RFUiScript RT-qPCR sample preparation reagent enablesefficient cell lysis, RNA stabilization, and removal ofgenomic DNA for sensitive quantitative gene expressionanalysis without RNA purification. This unique reagentaccelerates and streamlines RT-qPCR analysis of culturedcells by eliminating the need to purify RNA. Reversetranscription and real-time PCR can be performed directlyfrom cell lysates. This reagent is ideal for rapid, highthroughput gene expression analysis, such as validationof siRNA-mediated gene knockdown.Order Info: Pg 131021010 Rapid protocol (5–10 min) efficiently stabilizes RNA eagent enables multiplex real-time PCR detection of upRto 4 targets from as few as 10 cellsFor More InformationWeb: www.bio-rad.com/iscript-rt-qpcrRequest or download bulletin: 5736CellsCollect forRT-qPCR304010 miniScript RT-qPCR sample preparation reagent provides rapid,yet sensitive gene expression results for high-, medium-, andlow-copy number gene targets. HeLa cells were treated withiScript RT-qPCR sample preparation reagent at 125 cells/μl and theexpression levels of three different genes were assessed. 18S rRNA(—), b-tubulin (—), and CMYC (—) expression levels were determined byperforming reverse transcription (iScript cDNA synthesis kit) and qPCR(iQ SYBR Green supermix) directly from cell lysate preparations.RFU, relative fluorescence units.iScript RT-qPCR sample preparation reagent method:10 minLysisBind tocolumn matrix4 Sample Preparation20Cycles ensitive detection of high-, medium-, and low-copySnumber gene targets directly from cell lysatesSpin10DNasetreatmentElute forRT-qPCR 30 minStandard method: 30 min
RNA, DNA, and Protein Sample PreparationDNA Kits and Reagentswww.bio-rad.com/nasampleprepDNA Kits and ReagentsDNA Isolation Kit Selection GuideKitDNAYieldPreparationTimeBacterial CultureDensity or cuum**and spinSilicamembraneStandardQuantum Prep 40 µg 15 min1–10 mlSpinplasmid miniprepDiatomaceousearthEnrichedor standardQuantum Prep 400 µg45 min20–40 mlSpinplasmid midiprepDiatomaceousearthEnrichedor standardAurum plasmid 20 µg 10 minUp to 12 OD ml*mini* (OD600 of undiluted culture) x (culture volume in ml) # of OD ml; 1 OD600 is equivalent to approximately 8 x 108 cells/ml.** Using the Aurum vacuum manifold.Aurum Plasmid Mini KitOrder Info: Pg 13See AlsoTransfection reagents:pages 283–284.The easy-to-use Aurum plasmid mini kit improves theefficiency and throughput of plasmid purifications with a simplebind-wash-elute protocol using silica membranes, all in lessthan 10 minutes. Spin- and vacuum-mediated protocols areavailable. The purified plasmid DNA can be immediately usedin any downstream molecular biology application.Nucleic acidamplification:pages 296–319.The Aurum plasmid mini kit delivers high yields ofreproducible plasmid DNA preparations for: Automated fluorescence-based sequencingRestriction digestionLigation and transformationTransfectionPCRFor More InformationWeb: www.bio-rad.com/dna-isolationRequest or download bulletin: 2664Quantum Prep Plasmid KitsOrder Info: Pg 13Quantum Prep Plasmid Miniprep KitThe Quantum Prep miniprep kit offers high quality andyield in an easy spin-based procedure that takes less than15 minutes from cell culture to purified plasmid. PlasmidDNA is recovered in water or TE for immediate use in alldownstream molecular biology applications.Quantum Prep Plasmid Miniprep KitQuantum Prep Plasmid Midiprep KitThe Quantum Prep plasmid midiprep kit uses a simplespin-column procedure that significantly reduces the timerequired to generate the large quantities of plasmid DNAneeded to support applications such as transfection,subcloning, and other enzymatic manipulations.For More InformationWeb: www.bio-rad.com/dna-isolationRequest or download bulletin: 2325Quantum Prep Plasmid Midiprep KitSample Preparation 5
RNA, DNA, and Protein Sample PreparationSample Cleanup and PurificationSee AlsoMolecular biologyand biotechnologygrade resins:page 36.www.bio-rad.com/nasampleprepChelex -Based ResinsOrder Info: Pg 13The two Chelex-based resins available are composedof paired iminodiacetate ions coupled to a styrenedivinylbenzene matrix, providing highly selective removal ofpolyvalent cations that may inhibit PCR.Chelex 100 Molecular Biology Grade ResinNuclease- and ligase inhibitor–free, this pipettable, smallscale resin is certified not to inhibit PCR and ensurescomplete removal of PCR inhibitors and metal ions.InstaGene MatrixInstaGene matrix is designed for rapid isolation of smallamounts of genomic DNA of sufficient purity for PCR inunder an hour. The specially formulated 6% w/v Chelexresin adsorbs cell lysis products that interfere with PCR,leaving genomic DNA template in the supernatant whereit is immediately available for PCR reactions.For More InformationWeb: www.bio-rad.com/dna-isolationRequest or download bulletin: 2074Sample Cleanup and PurificationBio-Rad offers a variety of sample preparation and cleanup columns; many are also suitable for DNA fragment purification.See AlsoAgarose gel system:page 217.Nucleic acidstandards:pages 137–138.Freeze ’N Squeeze DNA Gel Extraction Spin ColumnsFreeze ’N Squeeze spin columns offer a filtration-basedpurification method, which provides a quick and effectivealternative to chemical extraction and electroelution methods.The Freeze ’N Squeeze method uses centrifugation to draw50–23,000 bp DNA out of agarose gel slices that have beenquickly frozen and thawed. Features include: 12Order Info: Pg 13345671,000 bpNo solutions to prepare — save time and avoid usingtoxic chaotropic materials700600500400300200100DNA immediately available for PCR, subcloning, ligations,and sequencing reactionsLess than 1 minute of hands-on timeRecovery of DNAfragments usingFreeze ’N Squeezespin columns.Extracted fragmentswere rerun on aReadyAgarose 1% TAEgel. Lane 1, precisionmolecular mass ruler;lane 7, EZ Load 100 bpPCR molecular ruler.For More InformationWeb: www.bio-rad.com/dna-cleanupFreeze ’N Squeeze Method–20ºC for 5 minPurified DNACut out gel slice.6 Sample PreparationChop into pieces.Transfer to spin columnand freeze for 5 min.Spin for 3 min.Recover purified DNA.
RNA, DNA, and Protein Sample PreparationSample Cleanup and Purificationwww.bio-rad.com/nasampleprepPrepacked Spin ColumnsOrder Info: Pg 13Prepacked size exclusion spin columns allow easycleanup and purification of DNA and proteins from lowerMW contaminants. Bio-Spin , Micro Bio-Spin , and PCRKleen columns clean up DNA or protein samples quicklyand easily using size exclusion chromatography. Thesecolumns are available in multiple sizes and offer multipleMW exclusion limits to accommodate a variety of needs.Use the chart below to choose the column that bestmeets your needs.Prepacked Spin Column Selection GuideBio-Spin 6Micro Bio-Spin 6Bio-Spin 30Micro Bio-Spin 30PCR KleenPacked supportSpecial gradeBio-Gel P-6 gelSpecial gradeBio-Gel P-6 gelSpecial gradeBio-Gel P-30 gelSpecial gradeBio-Gel P-30 gelSpecial gradesize exclusion gelEquilibration buffer10 mM Tris, pH 7.4,or SSC buffer*10 mM Tris, pH 7.4,or SSC buffer*10 mM Tris, pH 7.4,or SSC buffer*10 mM Tris, pH 7.4,or SSC buffer*10 mM Tris, 1 mMEDTA, pH 7.0Desalting ofoligonucleotides 20 bases ———Labeling reactions:removal of unincorporatednucleotides 20 basesor bp from DNA—— —Removal of primers andprimer-dimers fromPCR products 200 bp———— Buffer exchange(restriction fragments,PCR products, enzymereactions, sequencingtemplates) — —DNA sequencingreaction mixture cleanup**—— —Riboprobe cleanup***——— —Desalting of antibody,enzyme, and proteinsolutions — —Purification of proteinsof molecular weight 6,000 ———Purification of proteinsof molecular weight 40,000—— —1.1 ml0.7 ml1.1 ml0.7 ml0.6 ml90% recovery of20 bases or bp,99% retentionof salts90% recovery of20 bases or bp,99% retentionof salts95% recovery of22 bases or bp,98% retentionof ddNTPs95% recovery of22 bases or bp,98% retentionof ddNTPs85% recovery of 700 bp, 95%retention of primersand primer-dimersMolecular weightexclusion limit,globular proteins6,0006,00040,00040,0008,000,000Sample volume50–100 µl10–75 µl50–100 µl10–75 µl25–100 µlCentrifuge typeSwinging bucketMicrocentrifugeSwinging yYesYesYesYesYesApplicationsBed volumeRetention and recovery* 150 mM NaCl, 17.5 mM sodium citrate, pH 7.0.** In Tris buffer.*** In RNase-free Tris buffer.Sample Preparation 7
RNA, DNA, and Protein Sample PreparationProtein ExtractionSee AlsoEmpty columns:pages 66–69.Bio-Gel P media:page 47.Bio-Spin,Micro Bio-Spin, andMini Bio-Spin columns:page 63.Prepacked Bio-Spin and Micro Bio-Spin ColumnsBio-Spin and Micro Bio-Spin columns clean up and removesalts, nucleotides, dye terminators, and small molecules fromDNA, RNA, and protein samples in 10 minutes. Filled withspecially sized Bio-Gel P gels, these columns are shippedfully hydrated in Tris or SSC buffer. They yield 95% recoveryof DNA 22 bp and allow sample loads of 10–100 µl. For saferiboprobe preparation use RNase-free Micro Bio-Spin P-30Tris spin columns.For More InformationWeb: www.bio-rad.com/dna-cleanupPCR Kleen Spin ColumnsPCR Kleen columns are prepacked spin columns forpurifying PCR products and other DNA molecules 200 bpdirectly from reaction mixtures. A simple 4-minute spineffectively removes salts, nucleotides, enzymes, primers,and primer-dimers. Purified DNA fragments are immediatelyavailable for secondary PCR, subcloning, restriction digests,ligations, sequencing, and other enzymatic manipulations.For More InformationWeb: www.bio-rad.com/dna-cleanupRequest or download bulletin: 2311www.bio-rad.com/nasampleprepEnd capEnd capReservoirReservoir3 cm2 cm working bed height5 cm3.7 cm working bed height0.8 ml bedvolume1.2 ml bed volumePorous 30 µmpolyethylene bedsupport retainsfine particlesLuer endfitting withsnap-off tipLuer end fittingwith snap-off tipPrepacked Bio-Spin and Micro Bio-Spin ColumnsPCR Kleen Spin ColumnsProtein ExtractionProtein extraction tools such as cell lysis and extraction kits as well as mini grinders are available for extracting proteinsfrom cultured cells and tissues.See AlsoMicroRotofor cell:page 205.Kits for Cell LysisOrder Info: Pg 14MicroRotofor Lysis KitsThese kits are suitable for sample preparation for analytical(IPG strips) or preparative (Rotofor and MicroRotofor cells)IEF. The kits are based on a chaotropic protein solubilizationbuffer (PSB), which contains nondetergent sulfobetaine201 (NDSB 201) along with urea, thiourea, and CHAPSfor particularly effective solubilization (Vuillard et al. 1995).Resulting samples can be used directly for IEF. Cell lysisand extraction protocols are tailored for mammalian, plant,yeast, or bacterial samples.ReadyPrep Mini GrindersFor grinding small biological samples for high recovery ofproteins (and nucleic acids), each mini grinder includes a1.5 ml grinding tube containing a grinding resin and amatching pestle. The grinding resin is a neutral abrasivematerial made of high tensile-strength microparticles that donot bind proteins or nucleic acids. ReadyPrep mini grindersare disposable and are nuclease- and protease-free. Theyare a component of the MicroRotofor lysis kit (mammal) andare also sold separately as a pack of 20. The mini grindertubes fit conveniently in most benchtop centrifuges.8 Sample PreparationFor More InformationWeb: www.bio-rad.com/proteinextraction
RNA, DNA, and Protein Sample n Sample Cleanup, Reduction, and AlkylationBio-Plex Cell Lysis KitThe Bio-Plex cell lysis kit has been developed specifically toprepare cell culture and tissue lysate samples for analysiswith Bio-Plex phosphoprotein and total target assays.This cell lysis kit can also be used to prepare cell lysatesfor western blot analysis. Its protein extraction procedureyields western blotting results similar to those generated byroutine cell lysis and protein extraction protocols.For More InformationWeb: www.bio-rad.com/bioplexcelllysisRequest or download bulletins: 3033 and 3034ReadyPrep Protein Extraction KitOrder Info: Pg 14The ReadyPrep protein extraction kit (total protein)provides a simple, rapid, and reproducible method forpreparation of total cellular protein extracts from a widevariety of biological samples. Use of this kit generatesprotein samples that can be applied directly to a variety ofapplications, including IEF and 2-D gel electrophoresis.For More InformationWeb: www.bio-rad.com/proteinextractionRequest or download bulletins: 3033 and 3034Protein Sample Cleanup, Reduction, and AlkylationGeneral purpose cleanup kits and columns are available for the removal of salts and other contaminants.Protein Sample Cleanup Kit Selection GuideKit Type andCatalog NumberApplicationsProcedureSalt RemovalReadyPrep 2-D cleanupkit (#163-2130, #163-2140)Reduction of streaking on 2-D gelsConcentration of dilute samplesPreparation Acceptable Number ofTimeSample Sources PrepsTCA-like precipitation to remove salts, 1 hrs s s sdetergents, lipids, phenolic compoundsMicro Bio-Spin 6Removal of salts and other contaminants Size exclusion chromatography5 min—columns (see page 63)5025, 100,1,000Removal of Other ContaminantsReadyPrep 2-D cleanupReduction of streaking on 2-D gelskits (#163-2130, #163-2140) Concentration of dilute samplesReduction and AlkylationTCA-like precipitation to removesalts, detergents, lipids, phenoliccompounds 1 hrs s s s50ReadyPrep reductionReduction of streaking on 2-D gelsalkylation kit (#163-2090)Improved resolution of basic proteinsReduction, then alkylation of sample 2 hrto remove disulfide bonds and preventtheir re-formations s s s100s Mammalian tissue or cells. s Plant leaves. s E. coli. s Yeast.For More InformationWeb: www.bio-rad.com/proteincleanupRequest or download bulletins: 2934 and 2961Sample Preparation 9
RNA, DNA, and Protein Sample PreparationProtein ein FractionationFractionation kits reduce sample complexity, helping to identify low-abundance proteins. Fractionation kits can besubdivided into three groups that fractionate based on cellular location of the proteins of interest, differential solubility,and protein charge. Instruments are available to separate proteins by size and charge.Protein Extraction by Cellular LocationFractionation by Cellular LocationReadyPrep protein extraction kits facilitate fractionationof proteins from different cellular locations such as themembrane, nucleus, or cytoplasm. eadyPrep protein extraction kit (cytoplasmic/Rnuclear) — prepares fractions enriched in cytoplasmic ornuclear proteins from eukaryotic samples eadyPrep protein extraction kit (membrane I) —Roffers a quick and effective protocol for isolating mostmembrane proteins. It does not require ultracentrifugationor preparation of density gradientsOrder Info: Pg 15Fractionation Using Differential Protein SolubilityThe ReadyPrep sequential extraction kit and the ReadyPrepprotein extraction kit (soluble/insoluble) both reduce samplecomplexity using differential solubilization. The two kitscan be used independently, or the rehydration/samplebuffer from the soluble/insoluble kit can be used with thesequential extraction kit to create a fourth fraction for evenbetter resolution. eadyPrep protein extraction kit (membrane II) —Roffers a protocol for isolating more complexmembrane proteins eadyPrep protein extraction kit (signal) —Rfor isolating proteins involved in intracellular membranetrafficking and signaling pathways. These includeproteins such as GPI-anchored proteins, caveolinand associated proteins, acylated tyrosine kinases,and G proteins eadyPrep sequential extraction kit — enables theRisolation of three different fractions of increasing solubility.These fractions are isolated sequentially, allowing thevisualization of proteins that might not otherwise be seen.Increasing solubilization strength is provided through theuse of stronger detergents for each subsequent fraction eadyPrep protein extraction kit (soluble/insoluble) —Ruses a single fractionation stepReadyPrep Kit Components and Related ProductsIndividual ReadyPrep kit components and related productssuch as reducing agents are also available. See orderinginformation on page 15.For More InformationWeb: www.bio-rad.com/proteinfractionationRequest or download bulletins: 2934 and 2961Aurum Ion Exchange KitsAurum ion exchange (AEX or CEX) mini columns allowselective purification of acidic or basic proteins, respectively.These easy-to-use kits selectively enrich either acidicor basic proteins and can be used with a variety ofstarting samples.For More InformationWeb: www.bio-rad.com/proteinfractionationRequest or download bulletin: 292810 Sample PreparationOrder Info: Pg 15
RNA, DNA, and Protein Sample PreparationProtein Sample Depletionwww.bio-rad.com/proteinsampleprepProtein Sample DepletionComplex samples often require depletion of high-abundance proteins to allow the detection of the low-abundance ones.Bio-Rad offers two different methodologies for protein depletion — the ProteoMiner protein enrichment system(which utilizes hexapeptide libraries) and Aurum serum and Affi-Gel Blue products (which utilize resins).ProteoMiner Protein Enrichment SystemOrder Info: Pg 15The ProteoMiner protein enrichment system is a novelsample preparation tool for reducing the dynamic range ofprotein concentrations in complex biological samples. TheProteoMiner system: Enriches and concentrates low-abundance proteins thatcannot be detected through traditional methods orks with a variety of sample types (serum, plasma, urine,Wbile, cell lysates, tissues, and platelets) and is not limited bythe species’ origin tilizes a combinatorial library of hexapeptides ratherUthan immunodepletion, minimizing the dependence onavailable antibodies and preventing the codepletion oflow-abundance proteinsProteoMiner Protein Enrichment Kits — these kitscan be used with a variety of biological samples andare compatible with all major downstream proteomicsapplications. Small- and large-capacity kits are available forprocessing 2 or 10 samples.ProteoMiner Sequential Elution Kits — these kits utilizemultiple elution reagents to sequentially elute proteinsbased on different properties.ProteoMiner Small- and Large-Capacity KitsProteoMiner kits for protein enrichment are now offeredin formats optimized for varying starting amounts ofsample protein. Small-capacity kits — optimized for use with limitedsample material (minimum 10 mg of protein isrecommended) Large-capacity kits — optimized for use withsamples in which at least 50 mg of protein is availableFor More InformationWeb: www.bio-rad.com/proteominerRequest or download bulletins: 5632, 5635, and 5841Aurum Kits and ColumnsThe Aurum Affi-Gel Blue and Aurum serum kits andcolumns use affinity chromatography to reduce albumin andIgG, which improves analysis of lower abundance proteins.These products utilize a quick and easy spin-column formatand provide eluted proteins ready for analysis.For More InformationWeb: www.bio-rad.com/proteindepletionRequest or download bulletin: 2823Order Info: Pg minHeavychainIgGLightchainIgGRemoval of albumin and IgG from serum using the Aurum serumprotein mini kit. Total protein (1.32 mg) was purified on an Aurumserum protein mini column. Left, untreated serum; right, serum treatedwith the Aurum kit.Sample Preparation 11
RNA, DNA, and Protein Sample PreparationSurface-Enhanced Laser rface-Enhanced Laser Desorption/IonizationThe biomarker discovery process requires the analysis of large numbers of samples. The ProteinChip SELDI system’s highthroughput profiling technology allows you to easily acquire sufficient data for statistical significance, making the biomarkerdiscovery process more efficient.ProteinChip SELDI SystemSELDI combines the separation power of two techniques,chromatography and high-sensitivity mass spectrometry,which allows large numbers of proteins and peptides tobe detected and profiled. The ProteinChip SELDI systemprovides the arrays, reagents, kits, and software to rapidlygenerate a list of candidate disease biomarkers from largenumbers of samples.ProteinChip Arrays and Array PreparationProteinChip arrays utilize selective capture strategies toreduce sample complexity, allowing detection of lowabundance proteins for a number of applications, such asprotein profiling and biomarker discovery.ProteinChip KitsProteinChip kits contain all the reagents, consumables, andprotocols necessary to perform SELDI applications rangingfrom qualification of the ProteinChip SELDI reader throughprotein profiling and antibody capture.ProteinChip SoftwareSoftware applications tailored for the ProteinChipSELDI system enable fast, effective organization andanalysis of the large amounts of data generated duringbiomarker discovery.For More InformationWeb: www.bio-rad.com/proteinchipRequest or download bulletins: 5524 and 552612 Sample PreparationOrder Info: Pg 16
Ordering Informationwww.bio-rad.comRNA, DNA, and Protein Sample PreparationRNA, DNA, and Protein Sample PreparationRNA Kits and ReagentsCatalog #DescriptionAurum Total RNA KitsPg 3732-6820 Aurum Total RNA Mini Kit, 50 preps, includes 50 RNA binding columns, 50 capless collectiontubes (2.0 ml), 100 capped sample tubes (2.0 ml), 50 capped sample tubes (1.5 ml), 1 vial lyophilized DNase I,RNase-free reagents732-6830 Aurum Total RNA Fatty and Fibrous Tissue Kit, 50 preps, includes 732-6870, 50 ml PureZOL RNAisolation reagent732-6870* Aurum Total RNA Fatty and Fibrous Tissue Module, 50 preps, includes 50 RNA binding columns,50 capless collection tubes (2.0 ml), 100 capped sample tubes (2.0 ml), 50 capped sample tubes (1.5 ml),1 ml of lyophilized DNase I, RNase-free reagents and plasticware732-6800 Aurum Total RNA 96 Kit, 2 x 96-well preps, includes 2 grow blocks, sealing tape, 2 RNA bindingplates, 2 collection microplates, 2 vials lyophilized DNase I, RNase-free reagentsAurum Vacuum Manifold732-6470 Aurum Vacuum Manifold, for spin columns or 96-well plates, includes column adaptor plate, 4 replacementluer caps,
(iQ SYBR Green supermix) directly from cell lysate preparations. RFU, relative fluorescence units. iScript RT-qPCR sample preparation reagent method: Collect for 10 min Standard method: 30 min DNase treatment Lysis Spin Bind to column matrix RT-qPCR Elute for RT-qPCR 10 min 30 m
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Lab EX: Colony Morphology/Growth Patterns on Slants/ Growth Patterns in Broth (lecture only) - Optional Lab EX: Negative Stain (p. 46) Lab EX : Gram Stain - Lab One (p. 50) Quiz or Report - 20 points New reading assignment 11/03 F Lab EX : Gram Stain - Lab Two Lab EX: Endospore Stain (p. 56) Quiz or Report - 20 points New reading .
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