Nucleic Acid Purification Kits

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1 Nucleic Acid Purification Kits Fast Value Quality Convenient Reproducible Innovations in nucleic acid isolation

2 E.Z.N.A. Plasmid Mini Kit l For Isolation of up to 30 µg of plasmid DNA The E.Z.N.A. Plasmid Mini Kit I is designed to isolate up to 30 µg of high-quality plasmid DNA from 1-5 ml bacterial cultures in 30 minutes or less. This kit then uses a modified alkaline lysis method to lyse the cells and separate genomic DNA from plasmid DNA. Plasmid DNA purification is further simplified by using our HiBind mini-column technology in three quick steps: Bind, Wash, and Elute. Purified plasmid DNA is immediately ready for a wide variety of downstream applications such as routine screening, restriction enzyme digestion, DNA sequencing, cloning, transformation, and transfection. The E.Z.N.A. Plasmid Mini Kit I is available in two different column formats. V-Spin columns have an attached lid while Q-spin columns do not have a lid. Plasmid Mini Kit l Protocol Pellet Cells by Centrifugation Resuspend Lyse Neutralize Clear Lysate Transfer cleared lysate to HiBind Mini Column Q-Spin V- Spin Bind Wash 3x 300 Omega Bio-tek Plasmid Mini Kit l vs. Company Q Dry Elute Plasmid DNA in ng/µl ml DH5ἁ cultures transformed with pgem vector were grown for 24 hours in LB broth. Both protocols were performed according to manufacturer s recommended protocols. O.D. calculated using Thermo s NanoDrop 2000c. Total elution volume used was 50 µl.

3 E.Z.N.A. Plasmid Midi & Maxi Kit For isolation of Plasmid DNA from 50 or 200 ml Cultures The E.Z.N.A. Plasmid Midi Kit can isolate up to 200 µg from a 50 ml bacterial culture whereas the E.Z.N.A. Plasmid Maxi Kit can isolate up to 1 mg from a 200 ml culture. This kit then uses a modified alkaline lysis method to lyse the cells and separate genomic DNA from plasmid DNA. Cellular debri is removed by centrifugation and the cleared lysate is applied to a HiBind DNA column. Time consuming steps such as isopropanol precipitation are eliminated. Plasmid DNA is suitable for automated fluorescent DNA sequencing, restriction endonuclease digestion, transfection of mammalian cells, and many other downstream applications. E.Z.N.A. Plasmid Midi Kit Sample Culture Size A 260 /A 280 A 260 /A 230 Yield (µg) 1 50 ml ml ml ml E.Z.N.A. Plasmid Maxi Kit ml ml ml ml Bacterial cultures were grown for 24 hours in LB broth were used in respective culture amounts. DH5ἁ cells transformed with pgem vector. O.D. calculated using Thermo s NanoDrop 2000c. E.Z.N.A. Plasmid Midi Kit E.Z.N.A. Plasmid Maxi Kit

4 FastFilter Plasmid Midi & Maxi Kit Isolate Plasmid DNA in 40 Minutes FastFilter Plasmid DNA Kits Protocol Pellet bacterial cells by Centrifugation Resuspend Lyse Neutralize Add Binding Buffer Transfer Lysate into Lysate Clearance Syringe The Fastfilter Plasmid Kits combine the power of HiBind technology with the timetested consistency of alkaline-sds lysis of bacterial cells to deliver high-quality plasmid DNA. This system includes a special filter syringe, which replaces the centrifugation step following alkaline lysis. Plasmid DNA is suitable for automated fluorescent DNA sequencing, restriction endonuclease digestion, transfection of mammalian cells, and many other downstream applications. Insert plunger into Lysate Clearance Syringe & Clear Lysate Prepare Vacuum Manifold Insert column into luer connector on vacuum manifold Add Equilibration Buffer to DNA Column Pass Lysate through DNA Column Wash 3x Centrifuge to dry Elute by Centrifugation FastFilter Plasmid Midi Kit FastFilter Plasmid Maxi Kit

5 E.Z.N.A. Viral RNA Kit Isolate Viral DNA &RNA from Cell Free Fluid The E.Z.N.A. Viral RNA Kit is designed for the isolation of viral RNA and DNA from cellfree fluids such as plasma, serum, urine, and cell culture supernatant. The procedure completely removes contaminants and enzyme inhibitors, making viral RNA and DNA isolation fast, convenient, and reliable. This kit has been successfully used to isolate viral nucleic acids from hepatitis A, C, and HIV. The kit is also suitable for isolation of total RNA from cultured cells, tissues, and gram-negative bacteria. RNA purified using the E.Z.N.A. Viral RNA method is ready for applications such as RT-PCR and qrt-pcr E.Z.N.A. Blood DNA Kit Isolate DNA from Whole Blood The E.Z.N.A. Blood DNA Kit provides an easy and rapid method for the isolation of genomic, viral and mitochondrial DNA. Cellular DNA can be isolated from µl fresh or frozen whole blood. Omega Bio-tek s E.Z.N.A. Blood DNA Kit can also be used for the isolation of DNA from buffy coat, serum, plasma, saliva preserved in Oragene tubes, buccal swab and other body fluids. Sample Sample Size Average Yield (µg) Whole Blood 200 µl 4-8 Buffy Coat 200 µl Cultured Cells 1 x Buccal Swabs 1 Swab 2-4 Saliva 200 µl

6 E.Z.N.A. Cycle Pure Kit Purification of PCR Products Cycle Pure Protocol Add Buffer CP to PCR sample Apply CP/PCR to a HiBind Column Wash with DNA Wash Buffer The E.Z.N.A. Cycle Pure Kit is designed for the rapid purification of single or double stranded DNA from PCR or other enzymatic reactions. The purification procedure completely removes primers, primer-dimers, unicorporated nucleotides, enzymes such as polymerases, salts, and other impurities from the DNA sample. The E.Z.N.A. Cycle-Pure Kit takes less than 10 minutes to process from sample to purified DNA. Fragment sizes from 100 bp-10 Kb can be isolated with a recovery rate up to 85%. Purified DNA can be used in T-A cloning, sequencing, restriction enzyme digestion and various other labeling reactions. Dry Elute M Omega Bio-tek s Cycle Pure Kit vs. the Competition Company T Company A Company Q Company P Omega Bio-tek Company T Company A Company Q Company P M Omega Bio-tek Purified DNA from a 500 bp and 5 Kb PCR fragment was purified with Company T, Company A, Company P, Company Q, and Omega Bio-tek s Cycle Pure Kit and following the manufacturers recommended protocols. Ten percent of eluted DNA was analyzed on a 0.8% agarose gel. M: DL2000 Marker : PCR is a patented process of Hoffman-La Roche. Use of the PCR process requires a license.

7 E.Z.N.A. Gel Extraction Kit For Recovery of DNA Fragments from Agarose Gels Omega Bio-tek s E.Z.N.A. Gel Extraction kit uses HiBind DNA mini-column technology to recover DNA bands from 100 bp to 20 Kb in size. This kit can be used with all concentrations and types of agarose gels including TBE and TAE, with recovery rates up to 85%. The DNA band of interest is excised from the gel, dissolved in binding buffer, and applied to a DNA HiBind mini-column. Following three wash steps, DNA is eluted with elution buffer and is ready for downstream applications such as ligation, PCR amplification, restriction enzyme digestion, and various labeling reactions. The E.Z.N.A. Gel Extraction Kits can also be used to purify DNA fragments from PCR products and other enzymatic reactions. Recovery Rate of Excised DNA vs Company Q M C O Q C O Q C O Q C O Q M DNA ladder was run on a 2% agarose gel and four different fragment sizes (200 bp, 500 bp, 1Kb, and 5 Kb) were recovered using Omega Bio-tek s Gel Extraction Kit and Company Q kit for gel extraction following the manufacturer s recommended protocols. DNA was analyzed on a 2% TBE agarose gel with the respective companies eluate being compared to the original amount used in the gel extraction procedure. C: Control, O: Omega Bio-tek, Q: Company Q Control Omega Bio-tek Company Q Recovery Rate % bp 1000 bp 500 bp 200 bp Recovery percentage of E.Z.N.A. Gel extraction kit versus company Q comparable kit. The input amounts were normalized to 100%. O.D. readings taken with Thermo s Nanodrop C

8 E.Z.N.A. Tissue DNA Kit Isolate Genomic DNA from Cells and Tissue The E.Z.N.A. Tissue DNA Kit offers a simple, rapid and cost effective method for the isolation of DNA from a wide variety of sample sources including fresh or frozen tissues, animal cells, buccal swabs, and FFPE tissue sample. After cell lysis, the DNA purification process can be completed in less than 30 minutes. Up to 30 mg of tissue can be used, and multiple samples can be simultaneously processed with this spin-column based kit. There is no need for phenol/ chloroform extractions, or time consuming steps such as precipitation with isopropanol or ethanol. DNA purified using the E.Z.N.A. Tissue DNA Kit is ready for most downstream applications such as PCR, Southern blotting, microarrays, SYBR or probe based qpcr and restriction enzyme digestion. Omega Bio-tek s Tissue DNA Kit vs. the Competition DNA was isolated from 10 mg rat kidney using Omega Bio-tek s Tissue DNA kit. A Serial dilution of recovered DNA was then used in SYBR Green real time PCR targeting a 100 bp fragment of the GAPDH gene. Each reaction was performed in triplicate. The lines represent input DNA amounts of 10, 2, 0.4, 0.08, ng of DNA M Company T Company A Company P Company Q M Company M T Purified genomic DNA from 10 mg rat kidney tissue was isolated with Company T, Company A, Company P, Omega Bio-tek s Tissue DNA Kit and Company Q genomic DNA extraction kits following the manufacturers recommended protocols. Three percent of eluted DNA was analyzed on a 0.8% agarose gel. M: Lambda-Hind lll

9 E.Z.N.A. Total RNA Kit Isolate Total RNA from Cells and Tissue The E.Z.N.A. Total RNA Kit I provides a simple and rapid method for the isolation of up to 100 µg of total RNA from cultured eukaryotic cells and soft tissues. This kit enables simultaneous processing of multiple samples in less than 20 minutes. Typically up to 1 x 10 7 eukaryotic cells or 30 mg tissue can be used in a single experiment. Samples that can be used include but not limited to kidney, liver, heart, spleen, lung, pancreas, thymus, HeLa cells, HEK-293 cells, and HIN3T3 cells, Purified RNA can be used in many downstream applications such as RT-PCR, qrt-pcr, Northern blotting, Nuclease Protection Assay, microarrays and in vitro translation. Using the E.Z.N.A. Total RNA Kit l, 1µg of Total RNA was isolated from rat liver then reverse-transcripted with a M-MLV Kit. The cdna was diluted 0, 10, 100, 1,000 and 10,000 fold with water. Then used in a SYBR Green real time PCR reaction targeting a 100 bp region of the β-actin gene. Each reaction was performed in triplicates Source Sample Size RNA Yield (µg) Brain 10 mg 10 Kidney 10 mg 30 Liver 10 mg 45 Heart 10 mg 5 Spleen 10 mg 33 Lung 10 mg 12 Pancreas 10 mg 40 Thymus 10 mg 20 IC-21 1 x 10 6 Cells 12 HeLa 1 x 10 6 Cells 15 HEK x 10 6 Cells 10 HIN3T3 1 x 10 6 Cells 15 Expected RNA yields from mouse tissue samples and cultured cell types with the E.Z.N.A. Total RNA Kit I.

10 E.Z.N.A. SP Plant DNA Kit Isolate DNA from Plant Samples 100 mg of fresh leaf tissue Sample SP Plant DNA Company Q Yield (µg) Yield (µg) Sugar Cane Leaf Rice Leaf Arabidopsis Leaf Soy Bean Leaf Corn Leaf Tobacco Leaf The E.Z.N.A. SP Plant DNA Kit is specially designed for the rapid and reliable isolation of high-quality DNA from a variety of plant samples. Up to 100 mg fresh or 30 mg dry sample can be processed in less than 40 minutes. The optimized procedure incorporates the homogenizer spin column, a unique filtration and homogenization column that can efficiently remove debris and high molecular cellular components. Purified DNA is suitable for PCR, qpcr, restriction enzyme digestion, Southern blotting and hybridization applications. Tomato Leaf Peanut Leaf DNA was isolated from 100 mg peanut leaf using Omega Bio-tek s SP Plant DNA kit. A Serial dilution of recovered DNA was then used in SYBR Green real time PCR targeting a 91 bp fragment of the Alah1 gene. Each reaction was performed in triplicate. The lines represent input DNA amounts of 30, 3, 0.3, 0.03 ng of DNA

11 E.Z.N.A. Plant RNA Kit Isolate Total RNA from Plant Samples E.Z.N.A. Plant RNA Kit provides a convenient and rapid method for the isolation of total RNA from a variety of plant samples. This kit provides a homogenizer column for homogenization and filtration of viscous plant cell lysate in combination with HiBind RNA mini column technology for RNA purification. Contaminants such as polysaccharides and phenolic compounds are effectively removed. Purified RNA can be used for most downstream applications including RT- PCR, qrt-pcr, Northern blotting, differential display, micrroarrays, and poly A+ RNA selection. Omega Bio-tek s Yield vs. Company Q s Yield Company Q Lanes 1-3 Lanes Samples validated on E.Z.N.A. Plant RNA Kit Arabidopsis thaliana Humulus lupulus Beta vulgaris Fragaria virginiana Clarkia spp. Daucus carota Ornithogalum thyrsoides Dendranthema sp. Euglena gracilis Oryza sativa Hordeum vulgare Lycopersicon esculentum Malus sp. Solnm tuberosum Spinacia oleracea Surfinia sp. Triticum asestivum Vetis sp. Zea mays Nicotiana tabacum # Kit Yield (μg) A 260/ Company Q Company Q Company Q Total RNA was isolated from 100 mg Arabidopsis thaliana leaf tissue according to manufacturers recommended protocols. One tenth of eluate was analyzed on a denaturing agarose gel. O.D. values calculated with Thermo s Nanodrop 2000C

12 For Research Use. Only. Not for use in diagnostic procedures. SYBR is a registered trademark of Molecular Probes, Inc Nanodrop is a registered trademark of Thermo Fisher Scientific 0312