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I. Overview 1
II. Kit Components 1
III. Storage 2
IV. Intended Use 2
V. Safety Warnings and Precautions 2
VI. Warranty and Liability 2
VII. Technical Assistance 3
VIII. Quality Management 3
IX. Product Specifications 4
Poly A (Carrier RNA)
X. Principle 4
XI. Materials and Equipment Needed But Not Provided 5
XII. Protocols
Before You Begin 6
DNA Extraction from Forensic Sample 6
DNA Extraction from Small Amount of Tissue 10
DNA Extraction from Urine 11
DNA Extraction from Small Volumes of Blood and Saliva 12
DNA Clean-Up 13
XIII. Troubleshooting guide 15
XIV. Ordering information 17
XV. Explanation symbols 18
I. Overview
Description
MagListoTM 5M Forensic Sample DNA Extraction Kit utilizes Magnetic Nano Beads to extract
total DNA from a variety of forensic sample, such as whole blood, saliva, dried body fluid
TM
spot, fingerprint, nail clipping or hair using Magnetic Nano Beads and MagListo Magnetic
TM
Separation Rack. The use of MagListo Magnetic Separation Rack along with this kit
greatly increases user’s convenience by saving process time without use of centrifuge.
Applications
PCR, Real-Time PCR, SNP genotyping, STR (Short Tandem Repeat) analysis
Buffer ① (Lysis) 4 ml x 1 ea 40 ml x 1 ea
Buffer ② (Binding) 3 ml x 1 ea 30 ml x 1 ea
st
Buffer ③ (1 Washing) 4 ml x 1 ea 40 ml x 1 ea
nd
Buffer ④ (2 Washing) 1.6 ml x 1 ea 16 ml x 1 ea
Buffer ⑤ (Elution) 1 ml x 1 ea 15 ml x 1 ea
Proteinase K 5 mg x 1 ea 25 mg x 2 ea
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III. Storage
MagListoTM 5M Forensic Sample DNA Extraction Kit should be stored dry at room
TM
temperature. It can be stored for up to 1 year, if it remains sealed. MagListo 5M Forensic
Sample DNA Extraction Kit provides optimized Buffer ② (Binding) which is poisonous and
hazardous. Please, wear gloves and goggle eye protection when working with Buffer ②
(Binding).
Before, during and after use of this kit as described in this User’s Guide, all potentially
hazardous materials (i.e. materials that may have come in contact with genetically
recombinant samples) including tubes and tips should be processed and disposed of
according to applicable and appropriate regulations of the municipality/government in which
this product is being used. Users must be trained with basic experimental techniques for
correct execution of the experiments described in the User’s Guide.
Some applications that may be performed with this kit may infringe upon existing patents in
certain countries. The purchase of this kit does not include or provide a license to perform
patented applications. Users may be required to obtain a license depending on country and
application. We do not condone nor recommend unlicensed use of a patented application.
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compromise in product quality, immediately contact BIONEER’s Customer Service Center
(order@bioneer.com).
BIONEER does not assume liability for misuse of the product, i.e. usage of the product for
any purposes other than its intended purpose as described in the User’s Guide. BIONEER
assumes liability under the condition that users disclose all information related to the
problem in written form within 30 days of occurrence.
Technical Support
For all technical questions and troubleshooting on Bioneer products and applications.
Tel: +82-42-930-8777
Email: sales@bioneer.com
In North America
Tel: +1-877-264-4300
Email:support@bioneer.us.com
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IX. Product Specifications
Feature Specification
X. Principle
MagListoTM 5M Forensic Sample DNA Extraction Kit is designed for extraction of total DNA
from a variety of sources including high molecular weight up to 40 Kb (Note: this is
common for most DNA based application). Overall principle is based on adsorption of DNA
onto Magnetic Nano Beads by chaotrophic salt. For instance, chaotropic agent in Buffer ②
(Binding) contains guanidine hydrochloride. This removes water molecules around DNA and
silica coated Magnetic Nano Beads surface, resulting in total DNA being captured by
Magnetic Nano Beads. Magnetic Nano Beads and nucleic acid complexes are then pulled
and fixed on the tube wall using a magnetic force, followed by washing with ethanol to
remove debris and excessive salts. Captured nucleic acids are then eluted by Buffer ⑤
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(Elution), which contains an aqueous solution with optimal pH.
Please refer to the ordering information table on the latter part of the
manual, which contains the appropriate catalog number for specific tube
size.
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XII. Protocols
3. Buffer ② (Binding) contains chaotropic salt. You should take the appropriate laboratory
safety precautions and wear gloves when handling.
st nd
4. Add correct amount of absolute ethanol to Buffer ③ (1 Washing) and ④ (2 Washing).
※ Handling Samples
Dried body fluid spot or fingerprint (FTA card, paper, cloth etc.)
Punch out the sample up to 7 mm diameter using single-hole paper puncher or cut out up
2
to 2 cm . Cut the sample into smaller pieces to increase lysis efficiency.
Hair
Cut the hair 1 cm length from the hair root. Without root, cut up to 5 strands of whole hair
into small pieces.
Bone & teeth
Homogenize the bone up to 100mg into fine powder.
Chewing gum
Cut up to 30 mg of chewing gum into small pieces.
Cigarette butts
2
Cut out up to 2 cm piece of outer paper from the end of the cigarette butt.
Buccal swab
Cut the swab from its stick by hand or scissors. Use single piece of swab for extraction.
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2. (Lysis: 2-7) Add 300 μl of Buffer ⓛ (Lysis) and 10 μl of proteinase K solution (see
“Before you begin”) to the tube and completely resuspend the contents with vortex
mixer or pipetting.
3. (Optional) Add 20 μl of 1M DTT (Dithiothreitol, Not provided) to the tube and completely
resuspend the contents with vortex mixer. If the sample is hair, nail clipping or semen
stains, this step is necessary to increase sensitivity.
5. Add 300 μl of Buffer ② (Binding) to the tube and completely resuspend the contents by
vortex mixer or pipetting.
6. (Optional) Add 2 μl of dissolved poly A (see “Before you begin” or page 4 for details) to
the tube.
9. Take the clear supernatant only and transfer into new 1.5 ml or 2 ml tube.
10. (DNA Precipitation) Add 600 μl of absolute ethanol and mix well using a vortex mixer or
pipetting.
11. (DNA binding with Magnetic Nano Bead: 11-13) Add 100 μl of Magnetic Nano Bead
solution to the tube and mix thoroughly using a vortex mixer until Nano Beads are fully
resuspended.
(Note) Magnetic Nano Bead solution should be shaken well before each use.
TM
12. Place the tube in MagListo -2 Magnetic Separation Rack with magnet plate attached
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and invert the Rack with tube 3-4 times gently until Nano Beads bind tightly to magnet.
- Attachment
TM
13. Without removing the tube from MagListo rack, carefully pour supernatant out and
completely remove the remaining supernatant using a paper towel. In process,
magnetic crude pellet remains attached to the side of tube.
- Discard solution
Discard solution by inverting MagListo™ rack. Silicone immobilizer inside the stand holds
the tubes from falling in an upside down position. When discarding solution, invert rack
completely so the solution does not smear on the rack.
st TM
14. (1 washing: 14-16) Detach magnet plate from MagListo stand. Add 700 μl of Buffer
st
③ (1 Washing) to the tube. Close the cap and mix with vortex mixer until Nano Beads
are fully resuspended.
- Detachment
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TM
15. Attach the magnet plate to MagListo stand and invert the Rack with tube 3-4 times
gently until Nano Beads bind tightly to magnet.
TM
16. Without removing the tube from MagListo rack, pour supernatant out and remove
remaining supernatant using a paper towel by blotting.
nd nd
17. (2 washing) Repeat the above step 14 - 16 by adding 700 μl of Buffer ④ (2 Washing)
for additional washing.
rd
18. (3 washing) Repeat the above step 14 - 16 by adding 700 μl of absolute ethanol for
additional washing.
19. (Drying) Completely dry Nano Beads with the tube open and use a heat gun or a blow
dryer for 1 min 3 cm away from the top of the tube.
(Note) If you do not have using a heat gun or a blow dryer, place the rack lying down in
a dry oven at 60℃ for 10 min.
20. (Elution: 20-24) Add 30 - 100 μl of Buffer ⑤ (Elution) or distilled water to the tube with
magnet plate detached and resuspend the contents completely by pipetting or vortex
mixer for 15 sec.
TM
22. Attach magnet plate to MagListo stand and invert the Rack with tube 3-4 times gently
until Nano Beads bind tightly to the magnet.
TM
23. Without removing the tube from MagListo rack, carefully transfer supernatant
containing DNA to a sterile microcentrifuge tube.
24. Discard used Magnetic Nano Beads. Do not reuse Nano Beads.
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b. DNA Extraction from Small Amount of Tissue
3. Add 10 μl of proteinase K solution (see “Before you begin”) to the tube and mix
thoroughly using a vortex mixer.
6. Add 100 μl of Buffer ② (Binding) to the tube and mix immediately and thoroughly using
a vortex mixer. .
7. (DNA Precipitation) Add 200 μl of absolute ethanol and mix well using a vortex mixer or
pipetting.
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c. DNA Extraction from Urine
2. Add 500 ul of PBS and completely resuspend with vortex mixer or pipetting. Transfer the
sample to a 1.5 ml or 2 ml tube.
4. (Lysis: 4-8) Add 300 μl of Buffer ⓛ (Lysis) and 10 μl of proteinase K solution (see
“Before you begin”) to the tube and completely resuspend the contents with vortex
mixer or pipetting.
5. (Optional) Add 10 μl of 1M DTT to the tube and completely resuspend the contents with
vortex mixer. If the urine is expected to contain sperm cells, this step will improve yield.
7. Add 300 μl of Buffer ② (Binding) to the tube and completely resuspend by vortex mixer.
9. (DNA Precipitation) Add 600 μl of absolute ethanol and mix well using a vortex mixer or
pipetting.
10. Go to step 11 of “DNA Extraction from Forensic Sample” in page 7 and continue
extraction process.
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d. DNA Extraction from Small Volumes of Blood and Saliva
1. Add 10 μl of proteinase K solution (see “Before you begin”) to 1ml, 1.5 ml or 2 ml tube.
3. (Lysis: 3-4) Add 100 μl of Buffer ② (Binding) to each sample and mix immediately and
thoroughly using a vortex mixer. You must completely resuspend the sample to achieve
maximum lysis efficiency.
5. (DNA Precipitation) Add 200 μl of absolute ethanol and mix well using a vortex mixer or
pipetting.
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e. DNA Clean-Up
3. (Binding) Add 1 volume of Buffer ② (Binding) to the eluate and mix completely with
vortex mixer.
4. (DNA precipitation) Add 3 volumes of absolute ethanol to the eluate and mix well with
vortex mixer.
5. (DNA binding with Magnetic Nano Bead: 5-7) Add 100 μl of Magnetic Nano Bead
solution to the tube and mix thoroughly using a vortex mixer until Nano Beads are fully
resuspended.
(Note) Magnetic Nano Bead Solution contains magnetic nano beads. Please shake well
before use.
6. Place the tube in MagListo™-2 Magnetic Separation Rack with the magnet plate
attached and invert the Rack with tube 3~4 times gently until Nano Beads bind tightly to
magnet.
- Attachment
7. Without removing the tube from MagListo™ rack, carefully pour supernatant out and
completely remove remaining supernatant using paper towel by blotting.
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- Discard solution
Discard solution by inverting the MagListo™ rack. Silicone immobilizer inside the stand
holds the tubes from falling in an upside down position. When discarding solution,
invert rack completely so the solution does not smear on the rack.
st
8. (1 washing: 8-10) Detach the magnet plate from MagListo™ stand. Add 700 μl of
nd
Buffer ④ (2 Washing) to the tube. Close the cap and mix with vortex mixer until Nano
Beads are fully resuspended.
- Detachment
9. Attach magnet plate to MagListo™ stand and invert the tube 3~4 times gently until
beads bind tightly to magnet.
10. Without removing the tube from MagListo™ rack, pour supernatant out and remove
remaining supernatant using paper towel by blotting.
11. Go to step 18 of “DNA Extraction from Forensic Sample” in page 9 and continue clean-
up process.
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XIII. Troubleshooting Guide
st
Please check and add ethanol to the Buffer ③ (1 Washing) and ④
nd
(2 Washing). After adding ethanol, mix Washing Buffer well and
always mark the Washing Buffer bottles. If not, Washing Buffer
remain concentrated and may wash away the adsorbed DNA.
The lysis may have been incomplete. Please extend the incubation
Low yield of DNA
time. The amount of time for complete lysis varies depending on the
type of sample used and age of starting material. A shaking water
bath should be used for efficient lysis.
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Beads may have been dried insufficiently. You must completely dry
Nano Beads in a drying step. Remained ethanol can decrease purity
of DNA. Take enough time to completely dry Nano Beads.
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XIV. Ordering Information
K-3603 MagListoTM 5M Genomic DNA Extraction Kit, 100 reactions (mini) 1 kit
K-3604 MagListoTM 5M Plant Genomic DNA Extraction Kit, 8 reactions (mini) 1 kit
K-3605 MagListoTM 5M Plant Genomic DNA Extraction Kit, 100 reactions (mini) 1 kit
K-3610 MagListoTM 5M Cell Total RNA Extraction Kit, 8 reactions (mini) 1 kit
K-3611 MagListoTM 5M Cell Total RNA Extraction Kit, 100 reactions (mini) 1 kit
K-3614 MagListoTM 5M Forensic Sample DNA Extraction Kit, 8 reactions (mini) 1 kit
K-3615 MagListoTM 5M Forensic Sample DNA Extraction Kit, 100 reactions (mini) 1 kit
15 ml tube x 6
TM-1020 MagListoTM-15 Magnetic Separation Rack
holes
50 ml tube x 3
TM-1030 MagListoTM-50 Magnetic Separation Rack
holes
MagListoTM-2,
TM-1100 MagListoTM Magnetic Separation Rack Bundle Set -15, -50, and -96
(4 racks, 1 each)
K-3601-A Blow Dryer 1 ea
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XV. Explanation Symbols
Contains
Catalog Consult Instruction
sufficient for (n) USE BY
Number For Use
tests
Caution, consult
Temperature
Batch code accompanying Research Use Only
Limitation
documents
Caution,
DO NOT
Manufacturer Potential
REUSE
Biohazard
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