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MagListo 5M User Manual

Genomic dna extraction kit
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  • Page 2 Bioneer Corporation 8-11, Munpyeongseo-ro, Daedeok-gu, Daejeon 34302, Republic of Korea Tel: +82-42-930-8777 Fax: +82-42-930-8688 Email: sales@bioneer.co.kr www.bioneer.com...

  • Page 3: Table Of Contents

    Contents Overview Kit components Storage III. Intended use Safety warnings and precautions Warranty and liability Technical assistance VII. Quality management VIII. Product specifications Small Amount of Sample Principle Materials and Equipment Needed But Not Provided Protocols XII. Before You Begin DNA Extraction from Whole Blood DNA Extraction from Cultured Cell DNA Extraction from Animal Tissue...

  • Page 4: Overview

    I. Overview Description MagListo 5M Genomic DNA Extraction Kit utilizes the magnetic bead approach to extract total DNA from a variety of sources, such as whole blood, animal tissue and cultured cell using Magnetic Nano Beads and MagListo Magnetic Separation Rack. The use of...

  • Page 5: Storage

    Washing). Intended Use MagListo ™ 5M Genomic DNA Extraction Kit is intended for research use only. This kit is not intended for human or veterinary diagnostics. V. Safety Warnings and Precautions Please inquire BIONEER’s Customer Service Center to obtain a copy of the Material Safety Data Sheet (MSDS) for this product.

  • Page 6: Technical Assistance

    Every aspect of our quality management system from product development, production to quality MagListo assurance and supplier qualification meets the world-class standards. Each lot of ™ 5M Genomic DNA Extraction Kit is carefully tested by the quality control team. BQ-042-101-05 Revision : 2 (2016-07-04) www.bioneer.com...

  • Page 7: Product Specifications

    Small amount of sample MagListo 5M Genomic DNA Extraction Kit is also able to extract genomic DNA from a small quantity of sample. If the sample is low copy cell (< 1x10 ) or has a small amount of DNA, we recommend that about 4 μg of carrier DNA (a homopolymer such as poly-dA, poly-dT, or gDNA) or RNA should be added to the...

  • Page 8: Materials And Equipment Needed But Not Provided

    1. 1.5 ml or 2 ml tube, 15 ml tube, 50 ml tube 2. Vortex mixer 3. Absolute ethanol 4. Thermal block or dry oven 5. Phosphate buffered saline (PBS) 6. Blow dryer or heat gun MagListo Magnetic Separation Rack Magnetic Separation Rack Choice Tube MagListo Magnetic Separation Rack...

  • Page 9: Protocols

    XII. Protocols Before you begin 1. Completely dissolve Proteinase K (KB-0111) in 1,250 ul of nuclease-free water. Dissolved Proteinase K should be stored at 4℃. 2. Completely dissolve RNase A (KB-3101) in 600 ul of nuclease-free water. Dissolved RNase A should be stored at 4℃.
  • Page 10 Nano Bead solution to the tube and mix thoroughly using a vortex mixer until the beads are fully resuspended. (Note) Magnetic Nano Bead solution contains magnetic nano beads. Please shake well before use. 7. Place the tube in MagListo 2 (mini)/ MagListo 15 (midi)/...
  • Page 11 Detachment Push up the magnet plate gently. MagListo 10. Attach the magnet plate to rack and invert the tube 3 ~ 4 times gently until the beads tightly bind to the magnet. MagListo 11. Without removing the tube from rack, pour the supernatant out and remove the remaining supernatant using a paper towel by blotting.
  • Page 12 MagListo 18. Attach the magnet plate to rack and invert the tube 3 ~ 4 times gently until the beads tightly bind to the magnet. MagListo 19. Without removing the tube from rack, carefully take the supernatant containing DNA to a sterile microcentrifuge tube.

  • Page 13: Dna Extraction From Cultured Cell

    b. DNA Extraction from Cultured Cell for Micro/Mini/Midi/Maxi 1. Centrifuge the cultured cells (~ 1x10 (micro)/ ~ 1x10 (mini)/ ~ 5x10 (midi)/ ~ 1x10 (maxi)) for 10 min at 300 x g. Discard supernatant carefully. (Note) For cultured cells, “micro” scale is included to accommodate the use of lower cell number than 1x10 cells.
  • Page 14 9. Place the tube in MagListo 2 (micro, mini)/ MagListo 15 (midi, maxi) Magnetic Separation Rack with the magnet plate attached and invert the tube 3~4 times gently until the beads tightly bind to the magnet. Attachment Combine the magnet plate to the stand.
  • Page 15 MagListo 13. Without removing the tube from rack, pour the supernatant out and remove the remaining supernatant using a paper towel by blotting. washing) Repeat the above step 11 ~ 13 by adding 700 μl (micro, mini)/ 5 ml (midi)/ 10 ml (maxi) Buffer ④...
  • Page 16 Summary of Reagents Volume in Each Step of DNA Extraction from Cultured Cell Step Buffer Micro Mini Midi Maxi Page Cultured Cell ~ 1 x 10 ~ 1 x 10 ~ 5 x 10 ~1 x 10 P. 10 Lysis Buffer ②...

  • Page 17: Dna Extraction From Animal Tissue

    c. DNA Extraction from Animal Tissue for Mini/Midi/Maxi (Homogenization) Disrupt (or homogenize) the sample (~ 25 mg (mini)/ ~ 100 mg (midi)/ ~ 250 mg (maxi)) with a mortar and pestle. Place them to each specific tube format. a. (Mini) Place the homogenized tissue to a 1.5 ml or 2 ml tube. b.
  • Page 18 9. Place the tube in MagListo 2 (mini)/ MagListo 15 (midi)/ MagListo 50 (maxi) Magnetic Separation Rack with the magnet plate attached and invert the tube 3~4 times gently until the beads tightly bind to the magnet. Attachment Combine the magnet plate to the stand.
  • Page 19 12. Attach the magnet plate to MagListo stand and invert the tube 3 ~ 4 times gently until the beads tightly bind to the magnet. MagListo 13. Without removing the tube from rack, pour the supernatant out and remove the remaining supernatant using a paper towel by blotting.
  • Page 20 Summary of Reagents Volume in Each Step of DNA Extraction from Animal Tissue Step Buffer Mini Midi Maxi Page Animal tissue ~ 25 mg ~ 100 mg ~ 250 mg P. 14 Buffer ⓛ (Lysis) 180 μl 1.8 ml 3.6 ml P.

  • Page 21: Dna Extraction From Gram-Negative Bacteria

    d. DNA Extraction from Gram-Negative Bacteria for Mini/Midi/Maxi (Cell collection) Collect the bacteria cells (~1x10 (mini)/ ~5x10 (midi)/ ~1x10 (maxi)) by centrifugation at 6000 x g (> 8,000 rpm in a microcentrifuge) for 10 min (or >3,500 rpm for 20 min). And completely remove the media by pipetting.

  • Page 22: Dna Extraction From Gram-Positive Bacteria

    e. DNA Extraction from Gram-Positive Bacteria for Mini/Midi/Maxi (Cell collection) Collect the bacteria cells (~ 1x10 (mini)/ ~ 5x10 (midi)/ ~ 1x10 (maxi)) by centrifugation at 6,000 x (>8,000 rpm in a microcentrifuge) for 10 min (or >3,500 rpm for 20 min). And completely remove the media by pipetting.

  • Page 23: Dna Clean-Up

    Nano Bead solution to the tube and mix thoroughly using a vortex mixer until the beads are fully resuspended. (Note) Magnetic Nano Bead Solution contains magnetic nano beads. Please shake well before use. MagListo MagListo 6. Place the tube in ™...
  • Page 24 Discard solution Discard solution by inverting the MagListo ™ rack. The silicone immobilizer inside the stand holds the tubes from falling in an upside down position. When discard solution, invert the rack completely for the solution not to smear on the rack.

  • Page 25: Troubleshooting Guide

    XIII. Troubleshooting guide Comments and suggestions Buffers or other reagents may have been exposed to external changes or conditions that reduce its effectiveness. Please make sure that reagents were stored at room temperature at all times upon arrival and all reagent bottles were closed tightly, in order to preserve pH and stability, and to avoid contamination.
  • Page 26 MagListo ™ 5M Genomic DNA Extraction Kit copurify DNA and RNA when both are present in the sample. If RNA-free genomic DNA is required, RNase A Copurification of RNA should be added to the sample before addition of Buffer ② (Binding). If you want to remove RNA in the eluate, refer to “DNA Clean-Up”...

  • Page 27: Ordering Information

    1 kit MagListo K-3603 5M Genomic DNA Extraction Kit, 100 reactions (mini) 1 kit MagListo K-3604 5M Plant Genomic DNA Extraction Kit, 8 reactions (mini) 1 kit MagListo K-3605 5M Plant Genomic DNA Extraction Kit, 100 reactions (mini) 1 kit K-3606...

  • Page 28: Explanation Symbols

    XIV. Explanation Symbols Contains Catalog Consult Instruction sufficient for (n) USE BY Number For Use tests Caution, consult Temperature In Vitro Diagnostics Batch code accompanying Limitation Medical Device documents Authorized Caution, DO NOT Representative in Manufacturer Potential REUSE the European Biohazard Community BQ-042-101-05...

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