GE Healthcare Life Sciences illustra™ Nucleic Acid Sample Preparation Purify. Amplify. Simplify. Get attached with illustra nucleic acid purification illustra product range includes fast, efficient solutions for nucleic acid purification and amplification. From our illustra DNA sample prep kits to our illustra RNAspin kits for RNA purification, and illustra HotStart RTG™ and Master Mix for PCR, illustra delivers high-quality results with excellent purity and reproducibility. And illustra is easy to use, with simplified protocols that integrate colorcoded kit components, pre-equilibrated, columns, and minimal pipetting volume changes. Whatever your sample source, desired sample type, or downstream application, illustra makes nucleic acid purification and amplification a snap. www.vwr.com Selection guide for nucleic acid sample preparation Nucleic Acid Workflow Genome and Gene Analysis Sample(s): blood, tissue, cells, bacteria, yeast, etc. Plasmid DNA Preparation RNA Preparation Plasmid DNA Purification Plasmid Amplification Genomic DNA Preparation Genomic DNA Purification RNA Purification Whole Genome Amplification PCR RT-PCR PCR DNA Cleanup DNA Cleanup DNA Cleanup Cloning & Sequencing Gene Expression Genotyping or Cloning & Sequencing Biomolecule of Interest: e.g., Biomarker lasmid Genomic DNA, total RNA, and total denatured protein purification NEW illustra triplePrep™ Kit illustra triplePrep Kit is designed for rapid, simultaneous extraction and isolation of high quality genomic DNA (gDNA), total RNA and total dentured proteins from animal tissues and mammalian cells. High yields of quality DNA, RNA, and proteins can be extracted in less than 1 hour using a flexible, easy-to-follow workflow allowing researchers to directly correlate data generated from the same sample. The isolated DNA, total RNA, and total dentured proteins are suitable for genomic and proteomic applications such as PCR, restriction enzyme digestion, sequencing, array CGH, RT-PCR, gene expression microarray, SDS-PAGE, Western blotting, 2-D DIGE, and LCMS. Fast and simple. From sample to DNA/RNA/protein in less than 1 hour - streamlined workflow reduces the overall number of steps, resulting in up to 70% time saving compared to preparing each analyte individually. High yield. High DNA, RNA, and protein yields from small samples—optimized buffer, columns, and protocol ensure high recovery of gDNA, total RNA, and total denatured proteins. Easy to use • Flexible workflow. can isolate any two or all three analytes with multiple stop points in the protcol. •User friendly. minimal change of centrifugation speed, time and pipetting volume. High quality. • High purity—DNA-free RNA, with DNase provided in the kit. • Suitable for downstream applications—gDNA, RNA, and proteins validated in numerous downstream applications. •Easy to follow. color-coded caps and bottles with matching protocol steps minimize that chance for error. A quick refernce protocol card provides instructions at a glance for experienced users. Typical yields and purities from different tissues Tissue Liver Brain Heart Kidney Lung Spleen Tissue input amount (mg) Disruption difficulty Need extra spin 10 Easy No 10 Easy No 10 Hard No 10 Medium No 10 Medium Yes 5 Medium No DNA Yield (μg) Purity (A260/A280) 14.6 1.9 6.8 1.8 7.4 1.9 22.4 1.9 15.5 1.8 18.8 1.8 RNA Yield (μg) Purity (A260/A280) Need DNase 44 2.1 Yes 5.5 2.0 No 4.0 2.0 No 8.1 2.1 Yes 8.8 2.0 Yes 18.9 2.1 Yes Protein Yield (μg) 1460 746 923 897 592 510 Note: Yields and purities of gDNA, total RNA, and total denatured proteins can vary with user and depend on the nature and condition of the input sample. Typical yields and purities from different cells Cell Cell input amount (millions) 4 HeLa NIH-3T3 CHO-K1 HEK-293 1 1 1 1 DNA Yield (µg) Purity (A260/A280) 8.5 1.9 9.5 1.9 4.4 1.9 9.1 1.9 RNA Yield (µg) Purity (A260/A260) 13.7 2.1 7.9 2.1 6.6 2.0 10.1 2.0 Protein Yield (µg) 139 127 77 139 Ordering information Product Quantity VWR Cat. No. illustra triplePrep Kit 50 preps 95056-354 Whole genome amplification (WGA) illustra GenomiPhi™ DNA Amplification Kits Representative whole genome amplification (WGA) illustra GenomiPhi provides an easy way to prepare or replenish genomic DNA from limiting stocks or source materials. With as little as 10 ng of starting DNA, GenomiPhi can generate µg amounts of amplified DNA using a simple enzymatic reaction. Reactions are easily automated and scalable. Amplified DNA is suitable for use in a variety of downstream genotyping and cloning applications. illustra GenomiPhi V2 DNA Amplification Kit GenomiPhi V2 Quick mini-scale genomic DNA preparation: 4 - 7 μg in less than 2 hours. No background amplification generated in negative (no-template) control reactions. illustra GenomiPhi HY DNA Amplification Kit Midi-scale genomic DNA preparation: 40 - 50 μg in less than 4 hours. GenomiPhi HY GenomiPhi V2 7 GenomiPhi HY 6 60 10 ng gDNA NTC 10 ng gDNA NTC 50 4 40 Yield (µg) Yield (µg) 5 3 2 30 20 10 1 0 0 0 0.5 1 Amplification time (h) 1.5 2 0 1 2 3 4 Amplification time (h) Yield vs time plot for two GenomiPhi kits NTC: No template control; 10 ng DNA: 10ng purified genomic DNA was used as starting material. 5 Whole genome amplification (WGA) Success rate (%) illustra GenomiPhi DNA Amplification Kits GenomiPhiProtocol GenomiPhi 73 72 Input DNA (ng) 71 Sample buffer 70 69 68 GenomiPhi Number of wrong calls gDNA 6 5 4 3 2 1 0 REPLI-g ™ GenomePlex Heat or chemically denature. Reaction buffer Enzyme mix GenomiPhi REPLI-g GenomePlex Incubate at 30˚C. Inactivate enzyme at 65˚C. Representative amplification with illustra GenomiPhi DNA Amplification Kit Human genomic DNA samples were amplified in triplicate using various DNA amplification methods, followed by genotyping for over 5000 SNPs using the Linkage Panel IV chip performed by Illumina™ Genotyping Service. Amplified DNA produced with illustra GenomiPhi DNA Amplification Kit was 100% concordant with genomic DNA. Ordering information 6 Product (µg) Ordering information Product Quantity VWR Cat. No. Product Quantity VWR Cat. No. illustra GenomiPhi V2 DNA Amplification Kit 25 reactions 95042-266 illustra GenomiPhi HY DNA Amplification Kit 25 reactions 95042-268 illustra GenomiPhi V2 DNA Amplification Kit 100 reactions 95040-352 illustra GenomiPhi HY DNA Amplification Kit 100 reactions 95040-348 illustra GenomiPhi V2 DNA Amplification Kit 500 reactions 95040-354 illustra GenomiPhi HY DNA Amplification Kit 1000 reactions 95040-350 Plasmid DNA sequencing template preparation illustra TempliPhi™ DNA Amplification Kits Culture-free plasmid DNA preparation – straight from the plate Prepare DNA directly from bacterial colonies or glycerol stocks. Eliminates overnight cultures and conventional plasmid mini preps. Use amplified DNA directly in downstream DNA sequencing without further purification. illustra TempliPhi 100/500 DNA Amplification Kit • For low to medium throughput, small DNA vector preparation (e.g. pUC and M13). illustra TempliPhi 100 DNA Amplification Kit • High-quality DNA in as little as 4 hours. • Minimal hands-on time - easy to automate. illustra TempliPhi 2,000 Reaction Kit illustra TempliPhi Large Construct DNA Amplification Kit For higher-throughput sequencing template preparation. For large DNA vector preparation (e.g. fosmid and BAC). High quality DNA generates accurate sequencing results Plasmid DNA template was amplified with illustra TempliPhi 100 Amplification Kit and subsequently sequenced using DYEnamic™ ET Terminator Cycle Sequencing Kit and analyzed on an ABI™ PRISM™ 3100 Genetic Analyzer. Ordering information Product Quantity VWR Cat. No. illustra TempliPhi 100 reactions 89131-608 illustra TempliPhi 500 reactions 89131-612 illustra TempliPhi DNA Sequence Template Preparation Kit 2,000 reactions 89134-006 Plasmid DNA amplification using illustra TempliPhi DNA amplification kit 7 Difficult-to- sequence plasmid DNA templates illustra TempliPhi Sequence Resolver Kit Sequence resolution with a single kit — Solve the most common sequencing problems (repeats, sequencing stops, compressions) with one kit rather than multiple methods and reagents. Simple •Culture-less preparation—Amplify DNA from bacterial colonies, glycerol stocks, or purified DNA, saving costs in time, labor, and reagents. •Compatible with a variety of sequencing chemistries, including DYEnamic ET terminator and BigDye™ v3.1 terminator cycle sequencing kits. •Can be easily deployed upstream of standard sequencing protocols and instruments. High Quality Fast •More complete, accurate sequences—Improve difficult template sequencing success with up to 820 bp Phred 20 read lengths. •Just 15 min total hands-on time—Implement a simplified protocol with fewer steps and no overnight culturing. •No purification required—Use prepared DNA directly in sequencing reactions, eliminating purification steps to achieve higher sequencing throughput. Comparative resolution of GC-rich region with dGTP BigDye v3.0 and illustra TempliPhi Sequence Resolver Kit. Plasmid template (1 μl) was amplified using either illustra TempliPhi 100 Amplification Kit or illustra TempliPhi Sequence Resolver Kit. Amplified product (400 ng) was sequenced with either the dGTP BigDye kit (illustra TempliPhi 100 product) or the DYEnamic ET Terminator kit (TempliPhi Sequence Resolver product) per the standard protocols. Sequencing reactions were analyzed on an ABI 3730xl DNA Analyzer. Note the GC-rich region toward the end of the sequence is poorly resolved with the dGTP technique, resulting in miscalling. illustra TempliPhi Sequence Resolver Kit resolves compressions present with the dGTP technique. 8 Ordering information Product Quantity VWR Cat. No. illustra TempliPhi Sequence Resolver Kit 50 reactions 89131-616 illustra TempliPhi Sequence Resolver Kit 200 reactions 89131-618 Genomic DNA purification When your sample source is cell cultures or tissue, illustra tissue and cells genomicPrep™ kits provide fast extraction and purification of high-quality genomic DNA. illustra tissue and cells genomicPrep Mini Spin Kit allows you to go from sample to genomic DNA in just 90 min. illustra tissue and cells genomicPrep Midi Flow Kit reduces protocol time, including homogenization, by more than 2 h, as compared with other commercially available kits. illustra tissue and cells genomicPrep Mini Spin Kit Technical specifications For the rapid extraction and purification of high-quality genomic DNA from a variety of animal tissues and mammalian cell cultures. tissue & cells Mini Spin Kit (5 to 50 mg of Rat Liver; 20 mg optimum) Ordering information Yield Purity 12.0 to 25.0 μg 1.75 to 1.90 Total time of prep* 90 min Hands-on time* 20 min Product Quantity VWR Cat. No. Size illustra tissue and cells genomicPrep Mini Spin Kit 50 preps 95026-754 *prep time does not include homogenization illustra tissue and cells genomicPrep Mini Spin Kit 250 preps 20 to 35 kb Tissue & cells Mini Spin ( 3 × 105 - 5 × 106 cells) 95026-756 Yield 13 to 17 μg Purity 1.75 to 1.90 Total time of prep* Hands-on time* Size 70 min 20 min up to 75 kb *Cell type dependent, CHO and HK293 cells used in testing illustra tissue and cells genomicPrep Midi Flow Kit Technical specifications For the high-yield extraction and purification of high-quality genomic DNA from tissues and cells, 20% higher yield than other commercially available kits with an average size of > 20 kb. tissue & cells Midi Flow Kit (50 to 200 mg of tissue; 100 mg optimum) Ordering information Yield Purity 100 to 135 μg 1.7 to 1.9 Total time of prep* 3h Hands-on time* 1h Size Product Quantity VWR Cat. No. *prep time does not include homogenization illustra tissue and cells genomicPrep Midi Flow Kit 25 preps 95026-752 Tissue & cells Midi (1 × 107 to 2 × 107; 2 × 107 optimum) Yield Purity Total time of prep* Hands-on time* Size > 50 kb 100 to 135 μg 1.7 to 1.9 3h 1h > 50 kb *including lysis, purification & desalting 9 Genomic DNA purification illustra blood genomicPrep Mini Spin Kit For the rapid and reproducible isolation of high-quality genomic DNA from whole blood, buffy coat, bone marrow, and nucleated red blood cells. Ordering information Product Quantity VWR Cat. No. illustra blood genomicPrep Mini Spin Kit 50 preps 95026-740 illustra blood genomicPrep Mini Spin Kit 250 preps 95026-742 Technical specifications Yield 4 to 6 µg/200 µl whole blood Purity > 1.6 Total time of prep* less than 20 min Hands-on time* illustra blood genomicPrep Mini Spin Kit less than 5 min Size > 20 kbp *Including lysis, purification, and desalting illustra blood genomicPrep Midi Flow Kit For the rapid extraction and purification of high-quality genomic DNA from whole blood, buffy coat, and animal blood. Ordering information Product Quantity illustra blood 25 preps genomicPrep Midi Flow Kit VWR Cat. No. 95026-736 illustra bacteria genomicPrep Mini Spin Kit For the rapid extraction and purification of high-quality genomic DNA from Gram-positive and Gram-negative bacteria. Technical specifications blood Midi Flow Kit (1 to 8 ml; 5 ml optimum) Yield 85 to 120 μg Purity 1.7 to 1.9 Total time of prep* 2h Hands-on time* 45 min Size > 50 kb *including lysis, purification & desalting Technical specifications bacteria Mini Spin Kit (1 × 109 to 4 × 109 cells) Yield 4 to 12 μg Purity 1.7 to 1.9 Ordering information Total time of prep*- (Gram-negative) 40 min Product Quantity VWR Cat. No. Hands-on time* 15 min illustra bacteria genomicPrep Mini Spin Kit 50 preps 95026-732 Total time of prep*- (Gram-positive) 55 min illustra bacteria genomicPrep Mini Spin Kit 250 preps 10 95026-734 Hands-on time* 15 min Size > 20 kb *includes testing on a wide range of strains Genomic DNA purification Nucleon™ BACC Genomic DNA Extraction Kits Nucleon PhytoPure™ Genomic DNA Extraction Kits Developed for rapid extraction of high-quality, high-molecular- PhytoPure resin binds plant polysaccharides ensuring high weight genomic DNA from blood and cell cultures. yields of high-quality plant genomic DNA in only one hour. Ordering information Ordering information Product Quantity VWR Cat. No. Product Quantity VWR Cat. No. Nucleon BACC1 1 kit 95017-484 Nucleon PhytoPure 50 preps × 0.1 g 95026-702 Nucleon BACC2 1 kit 95026-698 Nucleon PhytoPure 50 preps × 1.0 g 95026-704 Nucleon BACC3 1 kit 95017-486 Nucleon HT Genomic DNA Extraction Kit Technical specifications Nucleon BACC1: 50 preps of 1 ml of whole blood or cultured cells (1 to 3 × 106 cells/ml) Nucleon BACC2: 50 preps of 10 ml of whole blood or cultured cells (3 to 10 × 106 cells/10 ml) or 100 preps of 1 ml (1 to 3 × 106 cells/ml) Nucleon BACC3: 50 preps of 10 ml of whole blood Facilitates the simple, phenol-free extraction of genomic DNA from paraffin-embedded sections and hard tissue requiring proteinase K digestion. Ordering information Product Quantity VWR Cat. No. Nucleon HT 1 kit, 50 preps 95026-700 Plasmid DNA purification illustra plasmidPrep Mini Spin Kit provides rapid isolation of highand low-copy number plasmid DNA for use in cloning, restriction enzyme digestion, PCR amplification, and DNA sequencing. For medium-scale cultures, illustra plasmidPrep Midi Flow Kit is ideal for transfection, sequencing, and enzymatic amplification and modification applications. illustra plasmidPrep Mini Spin Kit Technical specifications For the rapid isolation of high and low copy number plasmid DNA. plasmidPrep Mini (1.5 ml; A600 = 2.5 to 3.0) Yield Purity A260/A280 Purity A260/A230 Ordering information Product Quantity VWR Cat. No. illustra plasmidPrep Mini Spin 50 preps 95026-748 illustra plasmidPrep Mini Spin 250 preps 6 to 9 μg > 1.8 > 1.7 Total time of prep ~ 9 min Hands-on time ~ 3 min Size 6.3 kb plasmidPrep Mini (3.0 ml; A600 = 2.5 to 3.0) 95026-750 Yield Purity A260/A280 Purity A260/A230 9 to 15 μg > 1.8 > 1.7 Total time of prep ~ 9 min Hands-on time ~ 3 min Size 6.3 kb 11 Plasmid DNA purification illustra plasmidPrep Midi Flow Kit Technical specifications For the rapid extraction and purification of high-quality plasmid DNA from medium scale cultures for use in transfection, sequencing, and enzymatic amplification and modification. Ordering informationon Product Quantity VWR Cat. No. illustra plasmidPrep Midi Flow 25 preps 95026-744 Yield, purity, and quality values obtained with high- and low-copy number plasmid DNA purified with the plasmidPrep Midi Flow Kit. Plasmid copy number High Low Plasmid size (kb) 6.3 36 Culture volume (ml) 50 150 Overall plasmid DNA yield (μg) 289 191 Endotoxin (EU/μg pDNA) 0.05 0.13 A260/A280 ratio 1.90 1.93 A260/A230 ratio 2.34 2.29 Supercoiled momomer (%) 91.5 ND Transfection (% COS7 cells) 70 ND Sequencing (Phred20 read length) 818 ND plasmidPrep Midi (25 to 150 ml culture) Effective capacity Yield 250 μg 100 to 250 μg high-copy number plasmid; 20 to 250 μg low- and very-low-copy number plasmid % supercoiled pDNA endotoxin Total time of prep Hands-on time Size illustra plasmidPrep Midi Flow Kit Size Sephacryl™ S-1000 For large-scale purification of plasmid DNA by gel filtration. Ordering information Product Quantity VWR Cat. No. Sephacryl S-1000 SF 750 ml 95016-856 RNA purification Simple, convenient, and suitable for all levels of expertise, illustra RNAspin Mini Isolation Kit recovers high-quality total RNA from diverse sample types—even when your sample size is very small (e.g., 10 HeLa cells for RT-PCR). For applications requiring high yields, use illustra RNAspin Midi Isolation Kit with scalable input and output. illustra RNAspin 96 RNA Isolation Kit provides fast, efficient purification in a 96-well format, with flexible processing by vacuum or centrifugation. All illustra RNAspin kits can be used with cultured cells, tissue, bacteria, or yeast samples. 12 > 70% (strain dependent) < 10 EU per μg plasmid DNA ~ 215 min ~ 40 min 2 to > 50 kb RNA purification illustra RNAspin™ Mini Isolation Kit High-quality output RNA from diverse sample types; suitable for use in sensitive downstream applications. Ordering information Product Quantity VWR Cat. No. illustra RNAspin Mini Kit 20 preps 95017-489 illustra RNAspin Mini Kit 50 preps 95017-491 illustra RNAspin Mini Kit 250 preps 95017-493 Technical specifications Animal Tissue Cell Culture Sample size up to 30 mg tissue up to 5 × 106 cells Typical yield up to 70 μg up to 70 μg Elution volume 40–120 μl Effective binding capacity RNA integrity 100 μg sharp rRNA bands with no substantial degradative bands visible 28S:18S = ~ 2:1 RNA Integrity Number (RIN) values 7 RNA purity Total time of prep illustra RNAspin Mini Isolation Kit A260/A280 = 1.8–2.2 < 30 min/6 preps illustra RNAspin Midi Isolation Kit Technical specifications For applications requiring high yields. Ordering information Animal Tissue Cell Culture Sample size up to 200 mg tissue up to 5 × 107 cells Typical yield up to 700 μg up to 700 μg Product Quantity VWR Cat. No. Elution volume 500 μl illustra RNAspin Midi Kit 20 preps 95017-495 Effective binding capacity 700 μg RNA integrity sharp rRNA bands with no substantial degradative bands visible 28S:18S = ~ 2:1 RNA Integrity Number (RIN) values 7 A260/A280 = 1.8–2.2 RNA purity Total time of prep illustra RNAspin 96 RNA Isolation Kit Technical specifications For fast and efficient purification in a 96-well format. Ordering information Product 80 min/4 preps Animal Tissue Sample size Quantity VWR Cat. No. Typical yield Cell Culture Vacuum Centrifuge Vacuum Centrifuge 10–30 mg 30 mg tissue 2 × 106 1 × 107 up to 40 μg up to 100 μg up to 20 μg up to 100 μg illustra RNAspin 96 Kit 4 × 96 preps 95017-499 Elution volume illustra RNAspin 96 Kit 24 × 96 preps 95017-501 Effective binding capacity RNA integrity RNA purity Total time of prep 50–130 μl 100 μg sharp rRNA bands with no substantial degradative bands visible 28S:18S = ~ 2:1 RNA Integrity Number (RIN) values 7 A260/A280 = 1.8–2.2 70 min/96 preps 13 RNA purification illustra RNAspin 96 Filter Plates Allows for tissue samples to be processed with the RNAspin. illustra QuickPrep™ Micro mRNA Purification Kit/ illustra QuickPrep mRNA Purification Kit For rapid purification of mRNA directly from a wide variety of eukaryotic cells or tissue samples, even from a single cell. Ordering information Product Quantity VWR Cat. No. illustra RNAspin 96 Filter Plates 4 95026-706 illustra Cesium Trifluoroacetate (CsTFA) For separation and purification of nucleic acids, especially RNA, by isopycnic centrifugation. Ordering information Product Quantity VWR Cat. No. illustra QuickPrep Micro mRNA Purification Kit 1 kit 95017-636 illustra QuickPrep mRNA Purification Kit (4 purifications) 1 kit 95017-634 illustra mRNA Purification Kit Ordering information Product Quantity VWR Cat. No. illustra CsTFA (Solution) 100 ml 95016-998 For rapid affinity purification of mRNA from eukaryotic total RNA using prepacked oligo(dT)-cellulose spun columns. Ordering information Oligo(dT)-Cellulose Product Quantity VWR Cat. No. For simple, affinity-purification of polyadenylated mRNA. illustra mRNA Purification Kit 4 purifications (8 columns) 1 kit 95017-638 Ordering information Product Quantity VWR Cat. No. Oligo(dT)-Cellulose Type 7 1 g 95026-714 Oligo(dT)-Cellulose Type 7 5 g 95026-716 DNA Clean-up GE Healthcare provides a wide range of nucleic acid purification products for DNA clean-up. Whether your application needs require purification of PCR products and restriction fragments; removal of dye terminators, unincorporated labels and nucleotides; or desalting, illustra DNA clean-up products have been optimized for high performance in your specific application. The products have also been designed for convenience and speed with prepacked columns as well as quick reference protocol cards for ease of use. Many of the kits are tailored for a specific application to ensure purification of the highest quality DNA. illustra GFX™ PCR DNA and Gel Band Purification Kit Ordering information For the isolation and concentration of DNA fragments (50 bp to 10 kb) from PCR mixtures, DNA-containing agarose gel bands, enzyme-based DNA modifications, and restriction digestions. Product Quantity VWR Cat. No. illustra GFX PCR DNA and Gel Band Purification kit 100 purifications 95026-728 illustra GFX PCR DNA and Gel Band Purification kit 250 purifications 95026-730 14 DNA Clean-up illustra GFX 96 PCR Purification Kit For the purification of up to 96 PCR products (0.1 to 10 kb) simultaneously in as little as 15 min. Ordering information Product Quantity VWR Cat. No. illustra GFX 96 PCR Purification kit 10 × 96 well plates 95026-726 Technical specifications Protocol Elution volume (μl) Yield (%) Centrifugation 10 55 100 55 95 90 Vacuum 50 100 59 78 illustra GFX 96 PCR Purification Kit ExoSAP-IT™ illustra MicroSpin S-300 HR Columns For rapid and efficient purification of PCR products. For rapid purification of PCR products ( 100 bp) from unincorporated primers ( 20-mers) and nucleotides using spin-column chromatography. Ordering information Product Quantity VWR Cat. No. ExoSAP-IT 100 reactions 95026-708 Ordering information ExoSAP-IT 500 reactions 95026-710 Product Quantity VWR Cat. No. ExoSAP-IT 2000 reactions 95026-712 50 95017-617 ExoSAP-IT 5000 reactions 95040-358 illustra MicroSpin S-300 HR Columns illustra MicroSpin™ S-200 HR Columns For rapid purification of labeled single-stranded or doublestranded DNA fragments 50 bases in length in a volume of 51 to 100 μl using spin-column chromatography. Ordering information Product Quantity VWR Cat. No. illustra MicroSpin S-200 HR Columns 50 95017-615 illustra MicroSpin S-300 HR Columns illustra MicroSpin S-400 HR Columns For rapid purification of PCR products ( 200 bp) from unincorporated primers ( 32-mers) and nucleotides using spin-column chromatography. Ordering information Product Quantity VWR Cat. No. illustra MicroSpin S-400 HR Columns 50 95017-619 illustra MicroSpin S-200 HR Columns 15 DNA Clean-up Ordering information illustra MicroSpin G-25 Columns Product Quantity VWR Cat. No. illustra MicroSpin G-25 Columns 50 95017-621 Technical specifications Volume* % Average yield† Standard deviation Average purity‡ Standard deviation * † ‡ illustra MicroSpin G-25 Columns 100 μl 150 μl 83% 5.1 98% 2.9 89% 3.6 97% 4.8 A 17-mer was synthesized on a 0.2 μm scale and deprotected at 60°C for 30 min in concentrated ammonium hydroxide. 100 μl or 150 μl of deprotected oligo was applied to pre-spun MicroSpin G-25 Columns and centrifuged at 735 × g for 2 min in a microcentrifuge. As a standard, 0.25 ml of deprotected oligo was diluted to 1 ml and purified on a NAP-10 column according to instructions provided with the columns. Yield is based on A260 reading following purification. Yield from NAP-10 Column is a standard to which we have assigned 100% yield. Purity is based on amount of ammonia present following purification as determined using Nessler reagent. Purity from NAP-10 Column is a standard to which we have assigned 100% purity. illustra MicroSpin G-50 Columns illustra NICK™ Columns For rapid purification of labeled DNA ( 20 bases) from unincorporated labeled nucleotides in a volume of 25 to 50 μl using spin-column chromatography. For rapid purification of labeled DNA ( 20 bases in length) from unincorporated radiolabeled nucleotides using gravity-flow chromatography. Ordering information Ordering information Product Quantity VWR Cat. No. Product Quantity VWR Cat. No. illustra MicroSpin G-50 Columns 50 95017-623 illustra NICK Columns 20 95017-015 illustra MicroSpin G-50 Columns 250 95017-625 illustra NICK Columns 50 95017-017 illustra ProbeQuant™ G-50 Micro Columns illustra AutoSeq™ G-50 For rapid removal of unincorporated fluorescent dye-terminators from cycle sequencing reactions prior to automated sequencing. For rapid purification of labeled DNA ( 20 bases) from unincorporated labeled nucleotides in a volume of 25 to 50 μl using spin-column chromatography. Ordering information Ordering information Product Quantity VWR Cat. No. illustra AutoSeq G-50 50 columns 95017-629 illustra AutoSeq G-50 250 columns 95017-631 illustra AutoSeq G-50 1000 columns 95017-633 Product Quantity VWR Cat. No. illustra ProbeQuant G-50 Micro Columns 50 95026-724 illustra ProbeQuant G-50 Micro Columns illustra AutoSeq G-50 16 DNA Clean-up illustra NAP™ Columns Technical specifications For rapid and efficient purification of DNA and oligonucleotides (≥ 10-mers) in less than 15 min by gravity flow. Ordering informationrdering information Choosing a NAP column and buffer volumes Column Type Sample Volume (ml) Equilibration Buffer (ml) Elution Buffer (ml) NAP-5 0.1 0.4 0.5 Product Quantity VWR Cat. No. 0.25 0.25 0.7 illustra NAP-5 Columns 20 95017-007 Max. vol. 0.5 0 1.0 illustra NAP-5 Columns 50 95017-009 NAP-10 0.75 0.25 1.2 illustra NAP-10 Columns 20 95017-011 Max. vol. 1.0 0 1.5 illustra NAP-10 Columns 50 95017-013 NAP-25 1.5 1.0 2.5 illustra NAP-25 Columns 20 95017-003 2.0 0.5 3.0 2.5 0 3.5 illustra NAP-25 Columns 50 Max. vol. 95017-005 illustra Sephadex™ G-25 DNA Grade illustra CyScribe™ GFX Purification Kit For purification of DNA (≥ 10 bases in length) from small molecules by gel filtration. Allows rapid purification of CyDye™ labeled cDNA probes. Ordering information Ordering informationdering information Product illustra Sephadex G-25 DNA Grade SF Quantity 100 g Product illustra CyScribe™ GFX Purification Kit illustra CyScribe GFX Purification Kit VWR Cat. No. 95016-894 Quantity 25 purifications 50 purifications VWR Cat. No. 95026-722 95017-644 illustra Sephadex G-25 DNA Grade illustra Sephadex G-50 and G-100 DNA Grade For purification of DNA from small molecules by gel filtration. Ordering informationon Product illustra Sephadex G-50 DNA Grade F illustra Sephadex G-50 DNA Grade F illustra Sephadex G-50 DNA Grade M Quantity 25 g VWR Cat. No. 95016-896 100 g 95016-898 MicroSpin Columns 100 g 95016-782 Empty MicroSpin Columns for use with Sephadex DNA Grade media, or any media. illustra CyScribe GFX Purification Kit Ordering information Technical specifications G-25 DNA exclusion limit (base pairs) 10 Oligonucleotide exclusion limit [poly (A)] 10 ml of hydrated gel per gram of dry medium 4-6 G-50 G-100 20 20 9-11 25 27 15-20 Product MicroSpin Columns Quantity 100 VWR Cat. No. 95017-597 17 GE, imagination at work, and GE monogram are trademarks of General Electric Company. AutoSeq, CyDye, CyScribe, DYEnamic, genomicPrep, GenomiPhi, GFX, GraviTrap, GSTprep, GSTrap, HiTrap, HisTrap, HisPrep, illustra, LabMate, MBPTrap, MicroSpin, MidiTrap, MiniTrap, MultiTrap, NanoVue, NAP, NICK, Nucleon, PreScission, ProbeQuant, PuReTaq, QuickPrep, Ready-To-Go, RNA Spin, Sephacryl, Sephadex, Sepharose, SpinTrap, StrepTrap, triplePrep, TempliPhi, and Whatman are trademarks of GE Healthcare companies. CyDye: This product, or portions thereof, is manufactured under an exclusive license from Carnegie Mellon University under US patent number 5,268,486 and equivalent patents in the US and other countries. ExoSAP-IT: Exonuclease I/Shrimp Alkaline Phosphatase method of use is covered by one or more of the following US patent numbers: 5,756,285 and 5,741,676 in the name of GE Healthcare BioSciences Corp. Phi 29 DNA Polymerase: Phi 29 DNA polymerase and its use for DNA synthesis is covered by US patent numbers 5,854,033, 5,198,543, 5,576,204 and 5,001,050. Taq DNA Polymerase: This product is sold under licensing arrangements with Roche Molecular Systems, F Hoffmann-La Roche Ltd and the Perkin-Elmer Corporation. Purchase of this product is accompanied by a limited license to use it in the Polymerase Chain Reaction (PCR) process for research in conjunction with a thermal cycler whose use in the automated performance of the PCR process is covered by the upfront license fee, either by payment to Perkin-Elmer or as purchased (i.e., an authorized thermal cycler). FideliTaq is a trademark of USB Corporation. Illumina is a trademark of Illumina Inc. BigDye and PRISM are trademarks of Applera Corporation. GenomePlex is a trademark of Rubicon Genomics Inc. SEQUENOM and iPLEX are trademarks of Sequenom Inc. REPLI-g is a trademark of Qiagen. FideliTaq DNA Polymerase technology is licensed under U.S. Patent 5,436,149 owned by Takara Bio Inc. 2-D Protein Extraction Buffers, Mammalian Extraction Buffer and Yeast Extraction Buffer Kit are manufactured by g-Biosciences, St Louis, MO, USA.Shinjuku-ku, Tokyo 169-0073, Japan All third party trademarks are the property of their respective owners. © 2009-2010 General Electric Company - All rights reserved. First published May 2009. 1010 5M Lit. No. 07057REV 10/2010
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