Clean-up. Clean-up.

Transcription

1 DNA and RNA clean-up belong to the most frequent processes in molecular biology. MACHEREY-NAGEL s clean-up kits do not only focus on fast, easyto-handle, and quantitative high-recovery DNA and RNA purification, they also ensure high-quality downstream applications. Summary of products 40 Summary of procedures 41 PCR clean-up 44 Gel extraction 44, 50 Genomic DNA clean-up 51 RNA clean-up 53 Dye-terminator removal 55

2 and products Summary of products PCR clean-up Sample Product Page Single prep Mini spin columns (silica-membrane technology) Silica matrix (silica-matrix technology) < 400 μl PCR reaction mixture NucleoSpin Gel and PCR 44 < 400 μl PCR reaction mixture NucleoTraP CR 45 Manual and automated high throughput (HTP) 8-well strips, 96-well plates < 100 μl PCR reaction mixture NucleoSpin 8 PCR / Core Kit 46 (silica-membrane technology) NucleoSpin 96 PCR / Core Kit 96-well plates μl PCR reaction mixture NucleoFast 96 PCR 48 (ultrafiltration) Superparamagnetic beads (magnetic-bead technology) < 50 μl PCR reaction mixture NucleoMag 96 PCR 49 Gel extraction Sample Product Page Single prep Mini spin columns (silica-membrane technology) Silica matrix (silica-matrix technology) < 400 mg agarose gel NucleoSpin Gel and PCR 44 < 200 mg agarose gel NucleoTrap 50 Genomic DNA clean-up Sample Product Page Single prep (silica-membrane technology) Mini spin columns < 150 μl solution containing < 25 μg DNA Mini spin columns XS design < 400 μl solution containing < 2 μg DNA NucleoSpin gdna 51 NucleoSpin gdna XS 52 RNA clean-up Sample Product Page Single prep (silica-membrane technology) Mini spin columns < 200 μl phenol / chloroform extract or reaction mixture Mini spin columns XS design < 300 μl RNA solution containing < 90 μg RNA NucleoSpin RNA 53 NucleoSpin RNA XS 54 Dye-terminator removal Sample Product Page Single prep (gel filtration) Mini spin columns 20 μl sequencing reaction mixture NucleoSEQ

3 and products Summary of procedures PCR clean-up / Gel extraction PCR clean-up PCR clean-up / Gel extraction Single prep Manual and automated HTP Single prep NucleoSpin NucleoSpin NucleoSpin NucleoTraP CR / Gel and PCR 8 PCR 96 PCR NucleoTrap page 44 page 46 page 46 page 45 / 50 PCR sample agarose gel PCR sample PCR sample PCR sample NucleoTrap CR agarose gel NucleoTrap binding binding binding batch binding NucleoSpin PCR Binding Strips NucleoSpin PCR Binding Plate vacuum or centrifugation vacuum or centrifugation washing washing washing washing vacuum or centrifugation vacuum or centrifugation elution elution elution elution Silica-membrane technology Silica-matrix technology 41

4 and products Summary of procedures PCR clean-up Manual and automated HTP Dye-terminator removal Single prep NucleoFast NucleoMag NucleoSEQ 96 PCR 96 PCR page 48 page 49 page 55 hydration PCR sample PCR sample filtration binding column preparation NucleoFast 96 PCR Plate vacuum vacuum washing (optional) washing loading recovery elution recovery Ultrafiltration Magnetic-bead technology Gel filtration 42

5 and products Summary of procedures Genomic DNA clean-up RNA clean-up Single prep NucleoSpin NucleoSpin NucleoSpin NucleoSpin gdna gdna XS RNA RNA XS page 51 page 52 page 53 page 54 sample, e.g., pre-purified gdna, reaction mixtures sample, e.g., pre-purified gdna, reaction mixtures sample, e.g., pre-purified RNA, reaction mixtures, small cell numbers sample, e.g., pre-purified RNA, reaction mixtures binding binding binding binding washing washing washing washing elution in μl elution in 6 10 μl elution in μl elution in 5 30 μl Silica-membrane technology 43

6 PCR clean-up / Gel extraction NucleoSpin Gel and PCR Binding Buffer NTI colored ph indicator PCR clean-up and gel extraction the two-in-one kit with optimized recovery and elution volume Two applications, one kit one buffer with optimal performance for both applications High recoveries for fragments down to 50 bp Minimized elution volume of 15 μl highly concentrated DNA Binding buffer with ph indicator Separate buffers for single stranded DNA or SDS containing samples available Suitable for all gel buffer systems (e.g., TAE, TBE) NucleoSpin Gel and PCR Silica-membrane technology Mini spin columns < 400 μl PCR reaction mixture < 400 mg TAE / TBE agarose gel 50 bp approx. 20 kbp Typical recovery % A 260 /A Elution volume μl 10 min/6 preps Binding capacity 25 μg Procedure chart see page 41 Purification of PCR products Extraction of DNA from agarose gels DNA extraction from polyacrylamide gels of single stranded DNA and RNA extraction from agarose gels (Binding Buffer NTC required, not included in the kit, see ) of SDS-containing samples (Binding Buffer NTB required, not included in the kit, see ) Typical downstream applications: cloning, sequencing, PCR, restriction analysis NucleoSpin Gel and PCR 10 NucleoSpin Gel and PCR Columns, Collection Tubes (2 ml), buffers 50 as above as above Product accessories Pack of Specification REF Buffer NTB 150 ml for clean-up of SDS-containing samples Buffer NTC 100 ml for clean-up of single stranded DNA

7 PCR clean-up NucleoTraP CR The silica-matrix based kit for PCR clean-up without columns Scalable batch binding procedure Quantitative removal of primer and primer-dimer molecules High recovery for DNA fragments > 120 bp NucleoTraP CR Silica-matrix technology Silica bead suspension < 400 μl PCR reaction mixture 100 bp approx. 50 kbp Typical recovery % A 260 /A Elution volume μl 45 min/6 preps Binding capacity 0.6 μg/μl suspension Procedure chart see page 41 Purification of PCR products Concentration and desalting of reaction mixtures Typical downstream applications: cloning, sequencing, PCR, restriction analysis NucleoTraP CR 10 NucleoTraP CR Suspension, buffers as above

8 PCR clean-up NucleoSpin 8 PCR / Core Kit NucleoSpin 96 PCR / Core Kit of PCR products flexible 8-well strip format for varying throughput and proven 96-well plate format for high throughput Time-saving parallel clean-up of PCR products Complete removal of primers and primer-dimers optimal recovery Suitable for processing under vacuum and centrifugation Innovative MN Wash Plate minimizes risk of cross-contamination Suitable for manual or automated use NucleoSpin 8 / 96 PCR Core Kits: kits with basic content focusing on automation platforms. Additional accessories can be combined as needed. NucleoSpin 8 PCR NucleoSpin 8 PCR Core Kit NucleoSpin 96 PCR NucleoSpin 96 PCR Core Kit Silica-membrane technology Silica-membrane technology 8-well strips 96-well plates Processing Manual or automated, vacuum or centrifugation Manual or automated, vacuum or centrifugation < 100 μl PCR reaction mixture < 100 μl PCR reaction mixture 65 bp 10 kbp 65 bp 10 kbp Typical recovery % % A 260 /A Elution volume μl μl 30 min/6 strips 45 min/plate Binding capacity 15 μg 15 μg Procedure chart see page 41 Manual or automated purification of PCR products 46

9 PCR clean-up NucleoSpin 8 PCR 12 x 8 NucleoSpin PCR Binding Strips, MN Wash Plate, Rack of Tube Strips, buffers 60 x 8 as above NucleoSpin 8 PCR Core Kit 48 x 8 NucleoSpin PCR Binding Strips, buffers NucleoSpin 96 PCR 1 x 96 NucleoSpin PCR Binding Plate, MN Wash Plate, Elution Plate U-bottom, Self-adhering Foil, buffers 2 x 96 as above x 96 as above x 96 as above NucleoSpin 96 PCR Core Kit 4 x 96 NucleoSpin PCR Binding Plates, buffers 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 96 Vacuum Manifold Starter Set C 1 for use of NucleoSpin 8-well strips under centrifugation

10 PCR clean-up NucleoFast 96 PCR Cost-efficient 96-well ultrafiltration kit for PCR clean-up Ready-to-use DNA for sequencing and microarray spotting Sturdy membrane allows easy recovery of DNA No well-to-well cross-contamination Fast procedure Available as complete kits and as plates only Detergent-free membrane Manual or automated use NucleoFast 96 PCR Ultrafiltration 96-well plates Processing Manual or automated, vacuum or centrifugation μl reaction mixture > 150 bp Typical recovery % A 260 /A Recovery volume μl 20 min/plate (for typical PCR reactions of 25 μl) Procedure chart see page 42 Purification of PCR products > 150 bp NucleoFast 96 PCR Kit 4 x 96 NucleoFast 96 PCR Plates, Elution Plates U-bottom, Self-adhering Foil, buffers NucleoFast 96 PCR Plates 10 x 96 NucleoFast 96 PCR Plates x 96 as above Product accessories Pack of Specification REF NucleoFast Lids 50 lids for NucleoFast 96 PCR Plates NucleoVac 96 Vacuum Manifold NucleoVac Vacuum Regulator 1 for controlling of vacuum, recommended for use with NucleoVac 96 Vacuum Manifold 48

11 PCR clean-up NucleoMag 96 PCR Magnetic-bead based PCR clean-up Closed system reduced risk of cross-contamination Small elution volumes > 25 μl Recovery does not depend on elution volume Easily adaptable for automated use NucleoMag 96 PCR Magnetic-bead technology Highly reactive superparamagnetic beads Processing Manual or automated < 50 μl PCR reaction mixture 150 bp approx. 10 kbp Typical recovery % A 260 /A Elution volume μl min/96 preps Binding capacity 0.3 μg/μl beads Procedure chart see page 42 Manual or automated PCR clean-up Typical downstream application: sequencing NucleoMag 96 PCR 1 x 96 NucleoMag P-Beads, Elution Plate U-bottom, Self-adhering PE Foil, buffers 4 x 96 as above x 96 as above Material to be supplied by the user Pack of Specification REF Square-well Block (separation plate) 4 96-well blocks with 2.1 ml square wells as above Product accessories Pack of Specification REF NucleoMag SEP 1 magnetic separator

12 Gel extraction NucleoTrap The silica-matrix based kit for gel extraction without columns Scalable batch binding procedure High binding capacity even for very small fragments > 20 bp No shearing of large DNA fragments NucleoTrap Silica-matrix technology Silica bead suspension < 200 mg agarose gel 20 bp approx. 50 kbp Typical recovery % A 260 /A Elution volume μl 60 min/6 preps Binding capacity 0.6 μg/μl suspension Procedure chart see page 41 Extraction of DNA from agarose gels Concentration and desalting of reaction mixtures Typical downstream applications: cloning, sequencing, PCR, restriction analysis NucleoTrap 10 NucleoTrap Suspension, buffers as above NucleoTrap Suspension 100 NucleoTrap Suspension for 100 preps

13 Genomic DNA clean-up NucleoSpin gdna Post clean-up and concentration of large DNA fragments Effective post clean-up and concentration of large DNA fragments for successful downstream applications High recovery rates up to 90 % for DNA fragments 100 bp 50 kbp Concentrate your DNA sample easily and faster compared to microdialysis filtrations units 15 min/10 preps time-saving procedure NucleoSpin gdna Silica-membrane technology Mini spin columns < 150 μl aqueous DNA solution (< 25 μg DNA) 100 bp approx. 50 kbp Typical recovery % A 260 /A Elution volume μl 15 min/10 preps Binding capacity 50 μg Procedure chart see page 43 and concentration of pre-purified DNA (e.g., from organic extractions) and concentration of DNA from enzymatic reactions Not recommended for gel or PCR clean-up! For these applications, please use NucleoSpin Gel and PCR Cleanup (see page 44) Typical downstream applications: qpcr, enzymatic reactions, Southern blotting, STR amplification NucleoSpin gdna 10 NucleoSpin gdna Columns, Collection Tubes (2 ml), buffers 50 as above as above

14 Genomic DNA clean-up NucleoSpin gdna XS Post clean-up and concentration of small amounts of large DNA fragments Efficient post clean-up of large DNA fragments for successful downstream applications Recommended for small DNA quantities extracted from, e.g., forensic samples Unique XS spin column for low elution volume (> 6 μl) highly concentrated DNA Easier and faster than microdialysis filtration units 20 min/6 preps time-saving procedure NucleoSpin gdna XS Silica-membrane technology Mini spin columns XS design < 400 μl aqueous DNA solution (< 2 μg DNA) 100 bp approx. 50 kbp Typical recovery % A 260 /A Elution volume 6 10 μl 20 min/6 preps Binding capacity 3 μg Procedure chart see page 43 and concentration of pre-purified DNA (e.g., from organic extractions) and concentration of DNA from enzymatic reactions Not recommended for gel or PCR clean-up! For these applications, please use NucleoSpin Gel and PCR Cleanup (see page 44) Typical downstream applications: PCR, enzymatic reactions, Southern blotting, qpcr, STR amplification NucleoSpin gdna XS 10 NucleoSpin gdna XS Columns, Collection Tubes (2 ml), buffers 50 as above as above

15 RNA clean-up NucleoSpin RNA Simple, fast, and convenient clean-up of RNA Complete removal of RT-PCR inhibitors Time-saving procedure based on NucleoSpin RNA, without DNase digestion and homogenization steps RNA clean-up from pre-purified RNA (phenol / chloroform), or enzymatic reactions (e.g., amplification reactions, labeling reactions) NucleoSpin RNA Silica-membrane technology Mini spin columns < 200 μl phenol / chloroform extract, reaction mixture, or < 10 5 cells > 200 nt Typical recovery % A 260 /A Typical RIN (RNA integrity number) Depending on sample quality (no significant loss of RIN detected) Elution volume μl 20 min/6 preps Binding capacity 200 μg Procedure chart see page 43 * RNA clean-up of: Pre-purified RNA (e.g., TRIzol ) Reaction mixtures Amino-allyl-mRNA Biotinylated RNA RNA isolation from up to 10 5 cultured cells (whenever co-purification of some genomic DNA is acceptable, kit does not contain rdnase) Typical downstream applications: enzymatic labeling reactions, RT-PCR, DNA / RNA-based chip hybridizations * Kits to be used for research purposes only (see page 172) NucleoSpin RNA 10 NucleoSpin RNA Columns with Collection Tubes, Collection Tubes (2 ml), Collection Tubes (1.5 ml), buffers 50 as above as above

16 RNA clean-up NucleoSpin RNA XS Easy and fast concentration Highly efficient clean-up and concentration of RNA samples Complete removal of RT-PCR inhibitors Very high RNA recovery and concentration RNA clean-up from pre-purified RNA (phenol / chloroform), or enzymatic reactions (e.g., amplification reactions, labeling reactions, DNase digestions) Time-saving procedure based on NucleoSpin RNA XS technology Input volume up to 300 μl Elution in as little as 5 μl NucleoSpin RNA XS Silica-membrane technology Mini spin columns XS design < 300 μl RNA solution containing < 90 μg RNA > 200 nt Typical recovery % A 260 /A Typical RIN (RNA integrity number) Depending on sample quality (no significant loss of RIN detected) Elution volume 5 30 μl 20 min/6 preps Binding capacity 110 μg Procedure chart see page 43 * RNA clean-up of Pre-purified RNA (e.g., with TRIzol ) DNase digestions (e.g., with MACHEREY-NAGEL rdnase Set, page 155) Reaction mixtures Up to 55-fold increase in RNA concentration Typical downstream applications: enzymatic labeling reactions, RT-PCR, DNA / RNA-based chip hybridizations * Kits to be used for research purposes only (see page 172) NucleoSpin RNA XS 10 NucleoSpin RNA XS Columns with Collection Tubes, Collection Tubes (2 ml), Collection Tubes (1.5 ml), buffers 50 as above as above

17 Dye-terminator removal NucleoSEQ Pre-filled single spin columns for dye-terminator removal Efficient removal of dye terminators (e.g., BigDye terminators) without ethanol precipitation Convenient spin column format Long-term storage at room temperature NucleoSEQ Procedure chart see page 42 Dye-terminator removal Gel filtration Mini spin columns 20 μl sequencing reaction mixture 5 min/prep (without hydration of matrix) NucleoSEQ 10 pre-filled NucleoSEQ Columns, Collection Tubes (2 ml) as above as above Product accessories Pack of Specification REF Receiver Columns 20 μm 10 empty mini spin columns with 20 μm filter frit and Collection Tubes (2 ml), to be used for general filtration purposes as well as for retaining chromatographic resins 50 as above as above