RNA RNA.

Transcription

1 The world of nucleic acid analysis switches to the specific detection of different species from a variety of different starting materials. MACHEREY-NAGEL has become a new company offering a very broad range of innovative methods and products for sophisticated purification. The MACHEREY-NAGEL product line covers various applications, like simultaneous / DNA / protein isolation, ultra-sensitive extraction, or selective small purification. Summary of products 57 Summary of procedures 59 from cells and tissue 63 Micro 70, DNA, and protein isolation 72 from blood 76 Small and large from FFPE samples 80 from plant 82 Poly(A) m from total 83 clean-up (see Clean-up, page 53) / 56

2 Applications and products Summary of products from cells and tissue Sample Product Page Single prep (silica-membrane technology) XS design (silica-membrane technology) Midi spin columns (silica-membrane technology) Gravity-flow columns (anion-exchange chromatography) < 5 x 10 6 cultured cells, < 30 mg tissue NucleoSpin cultured cells, < 5 mg tissue NucleoSpin XS 64 < 5 x 10 7 cultured cells, < 200 mg tissue NucleoSpin Midi 65 < 2 x 10 7 cultured cells, < 100 mg tissue NucleoBond /DNA 66 Manual and automated high throughput (HTP) 8-well strips, 96-well plates < 10 7 cultured cells, < 20 mg tissue NucleoSpin 8 / Core Kit 67 (silica-membrane technology) NucleoSpin 96 / Core Kit Superparamagnetic beads (magnetic-bead technology) < 2 x 10 6 cultured cells, < 20 mg tissue NucleoMag Micro Sample Product Page Single prep (silica-membrane technology) < 10 7 cultured cells, < 30 mg tissue, < 50 mg plant tissue, < 150 μl reaction mixture < 300 μl plasma, serum (< 900 μl with multiple loading steps) NucleoSpin mi 70 NucleoSpin mi Plasma 71, DNA, and protein isolation Sample Product Page Single prep (silica-membrane technology), DNA, and protein isolation and protein isolation and DNA isolation Buffer set Protein quantification Buffer and reagent set < 5 x 10 6 cultured cells, < 30 mg tissue, < 100 mg plant tissue < 5 x 10 6 cultured cells, < 30 mg tissue, < 100 mg plant tissue < 5 x 10 6 cultured cells, < 30 mg tissue, < 100 mg plant tissue Protein dissolved in Protein Solving Buffer PSB or Laemmli buffer NucleoSpin TriPrep 72 NucleoSpin /Protein 73 NucleoSpin /DNA Buffer Set 74 Protein Quantification Assay 75 57

3 Applications and products Summary of products from blood Sample Product Page Single prep (silica-membrane technology) < 200 μl blood NucleoSpin Blood 76 (< 400 μl with two loading steps) Midi spin columns μl blood NucleoSpin Blood Midi 77 Manual and automated high throughput (HTP) 8-well strips, 96-well plates (silica-membrane technology) < 400 μl blood NucleoSpin 8 Blood NucleoSpin 96 Blood 78 Small and large from FFPE samples Sample Product Page Single prep (silica-membrane technology) 10 sections (10 μm) with < 50 mg tissue NucleoSpin total FFPE 80 XS design 10 sections (10 μm) with < 5 mg tissue NucleoSpin total FFPE XS 81 from plant Sample Product Page Single prep (silica-membrane technology) < 100 mg (wet weight), < 20 mg (dry weight) plant tissue NucleoSpin Plant 82 Poly(A) m isolation from total Sample Product Page Single prep (affinity chromatography) Latex beads < 250 μg total < 1000 μg total NucleoTrap m Mini NucleoTrap m Midi 83 clean-up (see Clean-up, page 53) Sample Product Page Single prep (silica-membrane technology) < 200 μl phenol / chloroform extract or reaction mixture XS design < 300 μl solution containing < 90 μg NucleoSpin Clean-up 53 NucleoSpin Clean-up XS 54 58

4 Applications and products Summary of procedures isolation NucleoSpin / NucleoSpin Plant page 63 / 82 sample, e.g., cells, tissue, bacteria, yeast, plant tissue lysis XS design NucleoSpin total FFPE page 80 NucleoSpin Blood page 76 FFPE sample whole blood paraffin removal lysis homogenization by filtration lysis NucleoSpin Filter NucleoSpin XS page 64 sample, e.g., small amounts of cells, tissue lysis homogenization by filtration (optional) NucleoSpin total FFPE XS page 81 FFPE sample paraffin removal lysis NucleoSpin Filter binding binding binding binding on-column rdnase digest on-column rdnase digest on-column rdnase digest (optional) on-column rdnase digest on-column rdnase digest (optional) elution in μl elution in μl elution in μl elution in 5 30 μl elution in 5 30 μl binding 59

5 Applications and products Summary of procedures isolation Midi spin columns NucleoSpin Midi page 65 sample, e.g., cells, tissue, bacteria, yeast Manual and automated HTP NucleoSpin NucleoSpin 8 / Blood Midi 8 Blood page 77 page 67 / 78 whole blood NucleoSpin 96 / 96 Blood page 67 / 78 NucleoMag 96 page 69 sample, e.g., cells, tissue, whole blood sample, e.g., cells, tissue lysis lysis lysis incl. homogenization homogenization by filtration homogenization by filtration (optional) NucleoSpin Filter Midi NucleoSpin Filter Strips (not included) lysis NucleoSpin Filter Plate (not included) binding binding binding NucleoSpin binding strips vacuum or centrifugation binding NucleoSpin binding plate vacuum or centrifugation on-column rdnase digest on-column rdnase digest on-column rdnase digest rdnase digest vacuum or centrifugation elution in μl elution in μl 60 elution in μl elution Magnetic-bead technology

6 Applications and products Summary of procedures Micro isolation NucleoSpin mi Plasma page 71 NucleoSpin mi page 70 sample, e.g., cells, tissue, plant plasma, serum lysis lysis homogenization by filtration protein precipitation NucleoSpin Filter binding of large binding (NucleoSpin Column) Purification of small and large in one fraction Purification of small and large in two separate fractions large bound to silica membrane small and protein in column flow-through protein precipitation on-column rdnase digest protein fraction large bound to silica membrane small and protein in column flow-through on-column rdnase digest removal of remaining protein elution of small and large protein fraction on-column rdnase digest (optional) removal of remaining protein (NucleoSpin Protein Removal Column) binding of small to column containing large protein precipitation (NucleoSpin Protein Removal Column) elution of large binding elution of small elution in μl 61

7 Applications and products Summary of procedures, DNA, and protein isolation NucleoSpin /Protein page 73 NucleoSpin TriPrep page 72 sample, e.g., cells, tissue, bacteria, yeast sample, e.g., cells, tissue, bacteria, yeast lysis lysis homogenization by filtration homogenization by filtration NucleoSpin Filter NucleoSpin Filter binding of / DNA / DNA bound to silica membrane protein purification / DNA bound to silica membrane purification protein in column flow-through protein purification / DNA purification protein in column flow-through binding of / DNA DNA DNA elution on-column rdnase digest of residual DNA elution in μl protein precipitation (special Protein Precipitator) of protein pellet redissolving pellet in PSB (Protein Solving Buffer) on-column rdnase digest elution in μl protein precipitation (special Protein Precipitator) of protein pellet redissolving pellet in PSB (Protein Solving Buffer) 62

8 from cells and tissue NucleoSpin Previously known as NucleoSpin II Mini spin kit for the isolation of of highest integrity rdnase included for on-column DNA digestion easy and efficient removal of contaminating DNA NucleoSpin Filters (shredders) efficient sample homogenization and reduction of viscosity Up to 70 μg ready-to-use Parallel purification of genomic DNA by using the NucleoSpin /DNA Buffer Set (page 74) NucleoSpin Technology < 5 x 10 6 cultured cells, < 10 9 bacterial cells, < 10 8 yeast cells, < 30 mg human / animal tissue Fragment size > 200 nt Typical yield 14 μg (10 6 HeLa cells), 70 μg (10 9 bacterial cells) A 260 /A Typical RIN ( integrity number) > 9 Elution volume μl Preparation time 35 min/6 preps Binding capacity 200 μg Procedure chart see page 59 Applications* isolation from cultured cells, tissue (standard protocol) from < 10 9 bacterial cells (Gram-negative, Gram-positive) or < 10 8 yeast cells from < 100 μl biological fluids clean-up from reaction mixtures from samples stored in later from saliva samples collected with Oragene (Genotek) Typical downstream applications: real-time RT-PCR, next generation sequencing, Northern blotting, primer extension, array technology, RNase protection assays * Kits to be used for research purposes only (see page 172) NucleoSpin 10 NucleoSpin Columns with Collection Tubes, NucleoSpin Filters, Collection Tubes (2 ml), Collection Tubes (1.5 ml), buffers, RNase-free rdnase 50 as above as above

9 from cells and tissue NucleoSpin XS Purification of highly concentrated from smallest samples Isolation of from small sample quantities like biopsy material or single cells Excellent recovery and integrity Concentrated for sensitive downstream applications by elution in as little as 5 μl rdnase included for on-column DNA removal NucleoSpin Filters (shredders) efficient homogenization and reduction of viscosity High quality, ready to use for RT-PCR and other applications Reducing Agent TCEP no β-mercaptoethanol necessary NucleoSpin XS Technology XS design cultured cells, < 5 mg human / animal tissue Fragment size > 200 nt Typical yield ng (10 2 HeLa cells) ng (10 4 HeLa cells) ng (10 3 HeLa cells) ng (10 5 HeLa cells) A 260 /A Typical RIN ( integrity number) > 9 (depending on sample quality) Elution volume 5 30 μl Preparation time 35 min/6 preps Binding capacity 110 μg Procedure chart see page 59 Applications* isolation from cultured cells isolation from tissue isolation from cryosections isolation from laser captured cells isolation from small amounts of plant material isolation from samples stored in later Typical downstream applications: real-time RT-PCR, next generation sequencing, Northern blotting, primer extension, array technology, RNase protection assays * Kits to be used for research purposes only (see page 172) NucleoSpin XS 10 NucleoSpin XS Columns with Collection Tubes, NucleoSpin Filters, Collection Tubes (2 ml), Collection Tubes (1.5 ml), buffers, RNase-free rdnase, Carrier, Reducing Agent TCEP 50 as above as above

10 from cells and tissue NucleoSpin Midi Previously known as NucleoSpin L Midi spin kit for the isolation of of highest integrity rdnase included for on-column digestion easy and efficient removal of genomic DNA NucleoSpin Filters Midi (shredders) efficient sample homogenization and reduction of viscosity Up to 600 μg ready-to-use NucleoSpin Midi Technology Midi spin columns < 5 x 10 7 cultured cells, < bacterial cells, < 3 x 10 8 yeast cells, < 200 mg human / animal tissue Fragment size > 200 nt Typical yield 180 μg (10 7 HeLa cells), 620 μg (4 x 10 7 HeLa cells) A 260 /A Typical RIN ( integrity number) > 9 Elution volume μl Preparation time 80 min/4 preps Binding capacity 700 μg Procedure chart see page 60 Applications* isolation from cultured cells, tissue (standard protocol) from < bacterial cells (Gram-negative, Gram-positive) or < 3 x 10 8 yeast cells clean-up from reaction mixtures from samples stored in later Typical downstream applications: real-time RT-PCR, next generation sequencing, Northern blotting, primer extension, array technology, RNase protection assays * Kits to be used for research purposes only (see page 172) NucleoSpin Midi 20 NucleoSpin Midi Columns with Collection Tubes, NucleoSpin Filters Midi with Collection Tubes, Collection Tubes (15 ml), buffers, RNase-free rdnase

11 from cells and tissue NucleoBond /DNA Anion-exchange chromatography extra high purity for up to 400 μg Ultra-pure from different samples Separate isolation of different species (t, r, m) Separate elution of genomic DNA NucleoBond /DNA 80 NucleoBond /DNA 400 Technology Anion-exchange chromatography Anion-exchange chromatography Midi gravity-flow columns Maxi gravity-flow columns 5 x 10 6 cultured cells, 20 mg human / animal tissue, 5 x 10 7 bacterial / yeast cells 2 x 10 7 cultured cells, 100 mg human / animal tissue, 2 x 10 9 bacterial / yeast cells Fragment size 50 nt 300 knt 50 nt 300 knt Typical yield 70 μg (5 x 10 6 cultured cells), 30 μg (20 mg tissue), 50 μg (5 x 10 7 bacterial cells) A 260 /A Preparation time h h Binding capacity 80 μg 400 μg 300 μg (2 x 10 7 cultured cells), 150 μg (100 mg tissue), 200 μg (2 x 10 9 bacterial cells) Applications* from cultured cells, tissue, bacterial cells * Kits to be used for research purposes only (see page 172) NucleoBond /DNA NucleoBond AXR 80 Columns, buffers NucleoBond /DNA NucleoBond AXR 400 Columns, buffers

12 from cells and tissue NucleoSpin 8 / Core Kit NucleoSpin 96 / Core Kit Isolation of flexible 8-well strip format for varying throughput and proven 96-well plate format for high throughput Time-saving parallel isolation of rdnase included for efficient removal of genomic DNA Processing under vacuum or by centrifugation Suitable for manual and automated processing Innovative MN Wash Plate minimizes risk of cross-contamination ready to use for any kind of enzymatic reaction NucleoSpin 8 / 96 Core Kits: Kits with basic content focusing on automation platforms. Additional accessories can be combined as needed. NucleoSpin 8 NucleoSpin 8 Core Kit NucleoSpin 96 NucleoSpin 96 Core Kit Technology 8-well strips 96-well plates Processing Manual or automated, vacuum or centrifugation Manual or automated, vacuum or centrifugation < 10 7 cultured cells (centrifugation), < 20 mg human / animal tissue (centrifugation), saliva (collected with Oragene ) Fragment size > 200 nt > 200 nt Typical yield 20 μg (2 x 10 6 HeLa cells), 20 μg (20 mg mouse liver) 20 μg (2 x 10 6 HeLa cells), 20 μg (20 mg mouse liver) A 260 /A Typical RIN ( integrity number) > 9 (cells) 7 (tissue) > 9 (cells) 7 (tissue) ratio 28S/18S ~ S/18S ~ 2.1 Typical concentration ng/μl ng/μl Elution volume μl μl Preparation time 45 min/6 strips 70 min/plate Binding capacity 100 μg 100 μg Procedure chart see page 60 Applications* Manual or automated isolation of from cultured cells and tissue from saliva samples collected with Oragene (Genotek) * Kits to be used for research purposes only (see page 172) For ordering information, please see next page 67

13 from cells and tissue NucleoSpin 8 12 x 8 NucleoSpin Binding Strips, MN Wash Plates, MN Square-well Blocks, Racks of Tube Strips, Elution Plate U-bottom, Self-adhering Foil, buffers, RNase-free rdnase 60 x 8 as above NucleoSpin 8 Core Kit 48 x 8 NucleoSpin Binding Strips, buffers, RNase-free rdnase NucleoSpin 96 2 x 96 NucleoSpin Binding Plates, MN Wash Plates, MN Square-well Blocks, Round-well Block Low, Elution Plates U-bottom, Self-adhering Foil, buffers, RNase-free rdnase 4 x 96 as above x 96 as above NucleoSpin 96 Core Kit 4 x 96 NucleoSpin Binding Plates, buffers, RNase-free rdnase Related products Pack of Specification REF NucleoSpin 96 Trace 2 x 96 for small sample amounts < 5 x 10 5 cultured cells, < 2 mg human / animal tissue NucleoSpin Trace Binding Plates, MN Wash Plates, Square-well Blocks, Elution Plates U-bottom, buffers, RNase-free rdnase 4 x 96 as above x 96 as above NucleoSpin 8 Tissue Core Kit 48 x 8 for larger tissue samples < 30 mg human / animal tissue NucleoSpin Tissue Binding Strips, buffers, RNase-free rdnase NucleoSpin 96 Tissue Core Kit 4 x 96 for larger tissue samples < 30 mg human / animal tissue NucleoSpin Tissue Binding Plates, buffers, RNase-free rdnase Product accessories Pack of Specification REF NucleoVac 96 Vacuum Manifold NucleoVac Vacuum Regulator 1 for controlling of vacuum Starter Set A 1 for use of NucleoSpin 8-well strips on the NucleoVac Vacuum Manifold Starter Set C 1 for use of NucleoSpin 8-well strips under centrifugation NucleoSpin Filter Plate 4 96-well plates for filtration of cell and tissue homogenates, for use under vacuum or centrifugation NucleoSpin Filter Strips 12 8-well strips for filtration of cell and tissue homogenates, F for use under vacuum or centrifugation 60 as above F 68

14 from cells and tissue NucleoMag 96 Magnetic-bead based isolation of from tissue and cell samples One-tube procedure minimizes risk of cross-contamination Small elution volumes > 50 μl Suitable for manual and automated processing Recombinant DNase included Reducing agent TCEP included (no β-mercaptoethanol necessary) Technology Processing Fragment size Typical yield Elution volume Preparation time Binding capacity Procedure chart see page 60 NucleoMag 96 Magnetic-bead technology Highly reactive superparamagnetic beads Manual or automated < 2 x 10 6 cultured cells, < 20 mg human / animal tissue > 200 nt < 30 μg μl 120 min/96 preps 0.3 μg/μl beads Applications* Rapid manual and automated small-scale preparation of highly pure from tissue or cell samples * Kits to be used for research purposes only (see page 172) NucleoMag 96 1 x 96 NucleoMag B-Beads, buffers, TCEP, RNase-free rdnase 4 x 96 as above Material to be supplied by the user Pack of Specification REF E.g., Rack of Tube Strips (lysis tubes) 4 sets 1 set consists of 1 rack, 12 strips with 8 tubes each, and 12 cap strips 24 sets as above Square-well Block (separation plate) 4 96-well blocks with 2.1 ml square wells as above E.g., Elution Plate U-bottom (elution plate) well microplates with 300 μl u-bottom wells Product accessories Pack of Specification REF NucleoMag SEP 1 magnetic separator KingFisher 96 Accessory Kit B (for use with KingFisher 96 platform) 1 set Square-well Blocks, Deep-well Tip Combs and Elution Plates for 4 x 96 preparations

15 Micro NucleoSpin mi No toxic phenol / chloroform convenient extraction Parallel isolation of small and large purification fractionated by size: Isolation of small only (< 200 nt), Isolation of small (< 200 nt) and large (> 200 nt) in two separate fractions Isolation of total (small and large in one fraction) Additional isolation of total protein fraction ready to use for SDS-PAGE and Western blot analysis Excellent recovery and purity by chaotropic salt lysis without phenol / chloroform (patent pending) NucleoSpin Filters for efficient sample homogenization rdnase for efficient on-column removal of genomic DNA Technology Fragment size Typical yield Elution volume Preparation time Binding capacity Procedure chart see page 61 NucleoSpin mi < 10 7 cultured cells, < 30 mg human / animal tissue, < 50 mg plant tissue, < 150 μl reaction mixture Small : < 200 nt, large : > 200 nt 10 μg small, 95 μg large (10 7 HeLa cells) μl 45 min/6 preps (small and large ) 35 min/6 preps (small only) 200 μg Applications* Parallel isolation of small and large from human / animal tissue and cultured cells, from plant tissue, and in combination with phenol / chloroform (e.g., TRIzol ) lysis Purification of si and large ds from DICER reactions Typical downstream applications: real-time RT-PCR, next generation sequencing, Northern blotting, chip hybridization * Kits to be used for research purposes only (see page 172) NucleoSpin mi 10** NucleoSpin Columns, NucleoSpin mi Columns, NucleoSpin Protein Removal Columns, NucleoSpin Filters, Collection Tubes (2 ml, 2 ml lid, 1.5 ml), buffers, RNase-free rdnase 50** as above ** as above ** Kits for 10 / 50 / 250 preps are sufficient for 10 / 50 / 250 preps when small and large is isolated in one fraction. When small and large is isolated separately, the kits are sufficient for 5 / 25 / 125 preps. 70

16 Micro NucleoSpin mi Plasma Isolation of small and DNA from plasma and serum Efficient and selective isolation of small from plasma or serum samples (patent pending) Superior recovery of small > 15 nt Optional on-column DNA digest for increased sensitivity in downstream applications Without phenol / chloroform simple and fast procedure Technology Fragment size Elution volume Preparation time Binding capacity Procedure chart see page 61 NucleoSpin mi Plasma 300 μl plasma or serum (up to 900 μl with multiple loading) < 1000 nt / bp μl 40 min/10 preps (without rdnase digestion) 70 min/10 preps (with rdnase digestion) 200 μg Applications* Isolation of small from plasma or serum for cancer research (e.g., reliable monitoring of clinical parameters) Typical downstream applications: real-time qrt-pcr, chip hybridization * Kits to be used for research purposes only (see page 172) NucleoSpin mi Plasma 10 NucleoSpin mi Columns, Collection Tubes (2 ml), buffers, RNase-free rdnase 50 as above as above

17 , DNA, and protein isolation NucleoSpin TriPrep Parallel isolation of, DNA, and protein from undivided samples Convenient one-column preparation of, DNA, and protein High quality and DNA, ready to use for downstream applications High protein yield independent of protein size, localization, modification, etc. Easy protein quantification using the MACHEREY-NAGEL Protein Quantification Assay (page 75) Complete kit including NucleoSpin Filters (shredders) for optimal lysis, rdnase for on-column DNA digestion, Protein Solving Buffer for solving all types of proteins NucleoSpin TriPrep Technology < 5 x 10 6 cultured cells, < 30 mg human / animal tissue, < 100 mg plant tissue DNA Protein Fragment size > 200 nt < 30 kbp kda Typical yield < 70 μg < 6 μg < 1200 μg A 260 /A Typical RIN ( integrity number) > 9 Elution volume ( and DNA) μl 100 μl μl Resolubilization volume (protein) Preparation time 30 min/6 preps () 45 min/6 preps ( + DNA) 35 min/6 preps (protein) Binding capacity 200 μg 10 μg* Procedure chart see page 62 * Binding capacity of DNA < 10 μg, strongly depending on amount bound to the membrane. Applications** Rapid purification of, DNA, and protein from small and precious samples no sample splitting for different isolations necessary Gene expression profiling, analysis of transgenic organisms, drug screening, genotyping Reliable interpretation of, DNA, and protein amounts Good preservation of protein primary structure and posttranslational modifications (e.g., protein phosphorylation) allows protein analysis without additional inhibitors (e.g., proteinase or phosphatase inhibitors) Suitable for a broad spectrum of starting materials, including cultured cells, tissue, bacteria, yeast, and plants and DNA are ready to use for all typical downstream applications Proteins are ready to use for SDS-PAGE / Western blotting ** Kits to be used for research purposes only (see page 172). DISTRIBUTION AND USE IN THE USA IS PROHIBITED FOR PATENT REASONS. NucleoSpin TriPrep 10 NucleoSpin TriPrep Columns with Collection Tubes, NucleoSpin Filters, Collection Tubes (2 ml), Collection Tubes (1.5 ml), buffers, RNase-free rdnase 50 as above as above

18 and protein isolation NucleoSpin /Protein Parallel isolation of and protein from undivided samples No splitting of samples required reliable interpretation of and protein extracted from the same sample (direct correlation) High yield and integrity High protein yield independent of protein size, localization, modification, etc. Easy protein quantification using the MACHEREY-NAGEL Protein Quantification Assay (page 75) Complete mini kit with NucleoSpin Filters (shredders) and recombinant DNase for DNA-free of high quality Technology NucleoSpin /Protein < 5 x 10 6 cultured cells, < 30 mg human / animal tissue, < 100 mg plant tissue Protein Fragment size > 200 nt kda Typical yield < 70 μg < 1200 μg A 260 /A Typical RIN ( integrity number) > 9 Elution volume () μl μl Resolubilization volume (protein) Preparation time 30 min/6 preps 35 min/6 preps Binding capacity 200 μg Procedure chart see page 62 Applications* Rapid purification of and protein from cultured cells and tissue Gene expression profiling, si experiments, analysis of transgenic organisms, drug screening Broad spectrum of starting material tested and proteins already detected * Kits to be used for research purposes only (see page 172) NucleoSpin /Protein 10 NucleoSpin /Protein Columns with Collection Tubes, NucleoSpin Filters, Collection Tubes (2 ml), Collection Tubes (1.5 ml), buffers, RNase-free rdnase 50 as above as above

19 and DNA isolation NucleoSpin /DNA Buffer Set Parallel isolation of and DNA from undivided samples in one working procedure Used in combination with NucleoSpin, NucleoSpin XS, NucleoSpin mi, NucleoSpin Blood, NucleoSpin Plant, NucleoSpin /Protein kits No need to split samples (e. g., precious, indivisible samples like biopsy material) High quality DNA and from one sample, perfect for PCR, RT-PCR, real-time PCR Fast procedure NucleoSpin /DNA Buffer Set Buffer set for elution of DNA in combination with preparations, to be used in combination with NucleoSpin kits See NucleoSpin, NucleoSpin XS, NucleoSpin mi, NucleoSpin Blood, NucleoSpin Plant, NucleoSpin /Protein DNA size < 30 kbp Typical DNA yield 5 μg (10 6 HeLa cells) 16 μg (30 mg pig liver) 5 μg (100 mg maize leaf) A 260 /A DNA elution volume 100 μl yield and purity Identical to NucleoSpin, NucleoSpin XS, NucleoSpin mi, NucleoSpin Blood, NucleoSpin Plant, NucleoSpin /Protein Applications* For biopsy material: for expression analysis; DNA for mutation analysis (e.g., of cancer genes) For ticks (Ixodes ricinus): for analysis of viruses (e.g., TBE-V, Tick-borne encephalitis virus, infectious agent of meningoencephalitis); DNA for analysis of germs (e.g., Borrelia burgdorferi, infective agent of the Lyme disease) For transgenic plants or animals: for expression analysis of the transformed gene and / or other genes; DNA for analysis of integration site, integration number and sequence confirmation of the transformed gene For transfected cultured cells: for expression analysis of the transgene or other genes of interest; DNA for analysis of methylation status of the transgene or other genes of interest * Kits to be used for research purposes only (see page 172). DISTRIBUTION AND USE IN THE USA IS PROHIBITED FOR PATENT REASONS. NucleoSpin /DNA Buffer Set 100 Buffers sufficient for 100 DNA isolations in combination with NucleoSpin, NucleoSpin XS, NucleoSpin mi, NucleoSpin Blood, NucleoSpin Plant, or NucleoSpin /Protein kits

20 Protein quantification Protein Quantification Assay Fast, sensitive, and convenient assay for protein quantification Highest sensitivity and flexibility Reducing agent and detergent compatible Fast procedure allows protein quantification in only 40 min The perfect supplement for NucleoSpin TriPrep, NucleoSpin /Protein, and NucleoSpin mi Reference protein (BSA) included Protein Quantification Assay Buffer and reagent set Sample size < 600 μl containing μg protein (BSA equivalents) Protein concentration Approx ng/μl Sample type Protein dissolved in Protein Solving Buffer PSB, Laemmli buffer, or equivalent, preferable free of nucleic acids Correlation cofficient Wavelength for light extinction measurement 570 nm ( nm) Assay time 40 min Applications Protein quantification assays in microplates microcuvettes semi-microcuvettes low-volume photometer (e.g., NanoDrop ) Protein quantification in Laemmli buffer Protein quantification in buffers containing up to 10 % SDS Protein quantification in buffers containing reducing agents (e.g., β-mercaptoethanol (715 mm), TCEP (50 mm), or DTT) Protein quantification in buffers containing bromophenol blue (e.g., 0.02 %) Protein Quantification Assay 50 Buffer PSB, Quantification Reagent, BSA as above

21 from blood NucleoSpin Blood Direct blood lysis no selective erythrocyte lysis Isolation of from fresh or frozen whole blood (human or animal) Direct total blood lysis very simple and convenient procedure (patent pending) No selective erythrocyte lysis at 4 C complete processing at room temperature Superior yield and quality from up to 400 μl whole blood Efficient on-column DNA removal for increased sensitivity in downstream applications Compatible with common blood collection tubes and anticoagulants (e.g., EDTA, citrate, and heparin) NucleoSpin Blood Technology < 400 μl whole blood (fresh or frozen) Fragment size > 200 nt Typical yield 1 8 μg (400 μl whole blood)* A 260 /A Elution volume μl Preparation time 55 min/6 preps Binding capacity 200 μg Procedure chart see page 59 * yield strongly depends on the leukocyte number in each individual blood sample. Applications** Isolation of from fresh or frozen whole blood from common blood collection tubes (EDTA, citrate, heparin) Typical downstream applications: qrt-pcr, next generation sequencing, blotting, array technology ** Kits to be used for research purposes only (see page 172) NucleoSpin Blood 10 NucleoSpin Blood Columns with Collection Tubes, Collection Tubes (2 ml), buffers, RNase-free rdnase, Liquid Proteinase K 50 as above

22 from blood NucleoSpin Blood Midi Direct blood lysis no selective erythrocyte lysis Midi spin kit for the isolation of from fresh or frozen blood (human or animal) Direct total blood lysis very simple and convenient procedure (patent pending) No selective erythrocyte lysis at 4 C complete processing at room temperature Superior yield and quality from up to 1.3 ml whole blood Efficient on-column DNA removal for an increased sensitivity in downstream applications Compatible with common blood collection tubes and anticoagulants (e.g., EDTA, citrate, and heparin) NucleoSpin Blood Midi Technology Midi spin columns μl whole blood (fresh or frozen) Fragment size > 200 nt Typical yield 4 26 μg (1300 μl whole blood)* A 260 /A Elution volume μl Preparation time 75 min/6 preps Binding capacity 700 μg Procedure chart see page 60 * yield strongly depends on the leukocyte number in each individual blood sample. Applications** Isolation of from fresh or frozen whole blood from common blood collection tubes (EDTA, citrate, heparin) Typical downstream applications: qrt-pcr, next generation sequencing, blotting, array technology ** Kits to be used for research purposes only (see page 172) NucleoSpin Blood Midi 20 NucleoSpin Blood Midi Columns with Collection Tubes, Collection Tubes (15 ml), buffers, RNase-free rdnase, Liquid Proteinase K

23 from blood NucleoSpin 8 Blood NucleoSpin 96 Blood Direct blood lysis no selective erythrocyte lysis Isolation of from whole blood (human or animal) flexible 8-well strip format for varying throughput and proven 96-well plate format for high throughput Direct total blood lysis very simple and convenient procedure (patent pending) No selective erythrocyte lysis at 4 C complete processing at room temperature Compatible with common blood collection tubes and anticoagulants (e.g., EDTA, citrate, and heparin) Suitable for fresh and frozen blood samples rdnase included for efficient removal of genomic DNA Processing under vacuum or by centrifugation Suitable for manual and automated processing Innovative MN Wash Plate minimizes risk of cross-contamination NucleoSpin 8 Blood NucleoSpin 96 Blood Technology 8-well strips 96-well plates Processing Manual or automated, vacuum or centrifugation Manual or automated, vacuum or centrifugation < 400 μl whole blood (fresh or frozen) < 400 μl whole blood (fresh or frozen) Fragment size > 200 nt > 200 nt Typical yield 1 8 μg 400 μl (whole blood)* 1 8 μg 400 μl (whole blood)* A 260 /A Elution volume μl μl Preparation time 60 min/6 strips 100 min/plate Binding capacity 100 μg 100 μg Procedure chart see page 60 * yield strongly depends on the leukocyte number in each individual blood sample. Applications** Manual or automated isolation of total from fresh or frozen whole blood stabilized with, for example, EDTA, citrate, or heparin Typical downstream applications: qrt-pcr, next generation sequencing, blotting, array technology ** Kits to be used for research purposes only (see page 172) 78

24 from blood NucleoSpin 8 Blood 12 x 8 NucleoSpin Blood Binding Strips, MN Wash Plates, Square-well Blocks, Racks of Tube Strips, Elution Plates U-bottom, Self-adhering Foil, buffers, RNase-free rdnase, Liquid Proteinase K 60 x 8 as above NucleoSpin 96 Blood 2 x 96 NucleoSpin Blood Binding Plates, MN Wash Plates, Square-well Blocks, Round-well Blocks Low, Elution Plates U-bottom, Self-adhering Foil, buffers, RNase-free rdnase, Liquid Proteinase K 4 x 96 as above Product accessories Pack of Specification REF NucleoVac 96 Vacuum Manifold NucleoVac Vacuum Regulator 1 for controlling of vacuum Starter Set A 1 for use of NucleoSpin 8-well strips on the NucleoVac Vacuum Manifold Starter Set C 1 for use of NucleoSpin 8-well strips under centrifugation

25 Small and large from FFPE samples NucleoSpin total FFPE Paraffin Dissolver no xylene required Two-in-one kit for the isolation of small (e.g., mi) and large from formalin-fixed, paraffin-embedded sample Blue colored Paraffin Dissolver (patent pending) included very easy and convenient paraffin removal rdnase included efficient on-column DNA removal High quality for enhanced RT-PCR performance Technology Typical yield Elution volume Preparation time Binding capacity Procedure chart see page 59 NucleoSpin total FFPE 10 sections (10 μm) with < 50 mg of tissue Depending on amount and quality of the sample μl 70 min/6 preps (90 min including optional rdnase digest) 200 μg Applications* Rapid isolation of from formalin-fixed, paraffin-embedded samples, e.g., colon carcinoma (unstained), colon carcinoma (hematoxylin stained), colon mucosa, adenocarcinoma of colon transversum, liver (rat / human), liver carcinoma (hematoxylin stained), lymph node, spleen (rat / human), pancreas (rat / human; diabetic / non-diabetic), placenta Isolation of from fresh and archived FFPE samples Isolation of from specimen on object slides (stained or unstained) Typical downstream application: RT-PCR * Kits to be used for research purposes only (see page 172) NucleoSpin total FFPE 10 NucleoSpin Columns, Collection Tubes (2 ml), Collection Tubes (1.5 ml), Paraffin Dissolver, buffers, Proteinase K, RNase-free rdnase 50 as above as above

26 Small and large from FFPE samples NucleoSpin total FFPE XS Improved yield with a new lysis chemistry Two-in-one kit for the isolation of small (e.g., mi) and large from even very limited formalin-fixed, paraffin-embedded samples Blue colored Paraffin Dissolver (patent pending) included very easy and convenient paraffin removal rdnase included efficient on-column DNA removal Higher sensitivity with concentrated elution volume as little as 5 μl High quality for enhanced RT-PCR performance Technology Typical yield Elution volume Preparation time Binding capacity Procedure chart see page 59 NucleoSpin total FFPE XS XS design 10 sections (10 μm) with < 5 mg of tissue Depending on amount and quality of the sample 5 30 μl 70 min/6 preps (90 min including optional rdnase digest) 100 μg Applications* Rapid isolation of and DNA, or only from formalin-fixed, paraffin-embedded samples, e.g., colon carcinoma (unstained), colon carcinoma (hematoxylin stained), colon mucosa, adenocarcinoma of colon transversum, liver (rat / human), liver carcinoma (hematoxylin stained), lymph node, spleen (rat / human), pancreas (rat / human; diabetic / non-diabetic), placenta Isolation of from fresh and archived FFPE samples Isolation of from specimen on object slides (stained or unstained) Typical downstream application: RT-PCR * Kits to be used for research purposes only (see page 172). NucleoSpin total FFPE XS 10 NucleoSpin FFPE XS Columns, Collection Tubes (2 ml), Collection Tubes (1.5 ml), Paraffin Dissolver, buffers, Proteinase K, RNase-free rdnase 50 as above as above

27 from plant NucleoSpin Plant Plant mini spin kit for all kinds of plant material Two alternative lysis buffers included optimized lysis procedure rdnase included for on-column digestion efficient removal of contaminating DNA NucleoSpin Filters (shredders) included efficient sample homogenization and reduction of viscosity Up to 70 μg ready-to-use Parallel purification of genomic DNA possible by using the NucleoSpin /DNA Buffer Set (page 74) NucleoSpin Plant Technology < 100 mg tissue Fragment size > 200 nt Typical yield 3 70 μg (100 mg plant material) A 260 /A Elution volume μl Preparation time 30 min/6 preps Binding capacity 200 μg Procedure chart see page 59 Applications* from plant cells and tissue from filamentous fungi Typical downstream applications: real-time RT-PCR, gene expression profiling, Northern blotting, primer extension, array technology, RNase protection assays * Kits to be used for research purposes only (see page 172) NucleoSpin Plant 10 NucleoSpin Plant Columns with Collection Tubes, NucleoSpin Filters, Collection Tubes (2 ml), Collection Tubes (1.5 ml), buffers, RNase-free rdnase 50 as above as above

28 Poly(A) m from total NucleoTrap m Mini NucleoTrap m Midi For fast purification of poly(a) m from total 5 μg poly(a) m/mg oligo(dt) latex beads (20 μl bead suspension) high binding capacity Direct m isolation from cells High quality poly(a) m without degradation and DNA contamination Convenient and fast processing by microcentrifugation using NucleoTrap Microfilters m purification in only 30 min Available in mini and midi format NucleoTrap m Mini NucleoTrap m Midi Technology Affinity chromatography Affinity chromatography Oligo(dT) latex-bead suspension Oligo(dT) latex-bead suspension < 250 μg total < 1000 μg total Fragment size 50 nt 20 knt 50 nt 20 knt Typical yield 10 μg m 40 μg m A 260 /A Elution volume μl μl Preparation time 30 min/6 preps 30 min/6 preps Binding capacity 0.25 μg poly(a) m/μl oligo(dt) latex-bead suspension Applications* Poly(A) m isolation from total Clean-up of in-vitro transcripts Direct purification of poly(a) m from cells * Kits to be used for research purposes only (see page 172) NucleoTrap m Mini 12 NucleoTrap Microfilters, Microcentrifuge Tubes (2 ml), Oligo(dT) Latex Beads, buffers NucleoTrap m Midi 12 NucleoTrap Microfilters, Microcentrifuge Tubes (2 ml), Oligo(dT) Latex Beads, buffers