MagListo 5M User manual

Bioneer Corporation
8-11, Munpyeongseo-ro, Daedeok-gu, Daejeon
34302, Republic of Korea
Tel: [phone]
Fax: [phone]
Email: [email protected]
www.bioneer.com

Contents
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
Small Amount of Sample
4
X.
Principle
4
XI.
Materials and Equipment Needed But Not Provided
5
XII.
Protocols
6
Before You Begin
6
DNA Extraction from Whole Blood
6
DNA Extraction from Cultured Cell
10
DNA Extraction from Animal Tissue
14
DNA Extraction from Gram-Negative Bacteria
18
DNA Extraction from Gram-Positive Bacteria
19
DNA Clean-Up
20
XIII.
Troubleshooting guide
22
XIV.
Ordering information
24
XV.
Explanation symbols
25

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I. Overview
Description
MagListo
TM 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
TM Magnetic Separation Rack. The use of
MagListo
TM Magnetic Separation Rack along with the kit
greatly increases user convenience by saving process time without the need of centrifuge.
Features and Benefits
- Magnetic Nano Beads enable rapid extraction.
- No costly capital instrument to instruments except MagListo
TM Magnetic Separation Rack.
Applications
Gene Cloning, PCR, Real-Time PCR, Southern Blotting, SNP genotyping
II. Kit Components
MagListo
TM 5M Genomic DNA Extraction Kit
Cat. no. 3602, 3603
*K-3602
(8 reaction)
**K-3603
(100 reaction)
Buffer ①(Lysis)
2 ml x 1 ea
25 ml x 1 ea
Buffer ②(Binding)
2 ml x 1 ea
25 ml x 1 ea
Buffer ③(1st Washing)
6 ml x 1 ea
60 ml x 1 ea
Buffer ④(2nd Washing)
1.6 ml x 1 ea
16 ml x 1 ea
Buffer ⑤(Elution)
1 ml x 1 ea
25 ml x 1 ea
Magnetic Nano Bead - DNA
1 ml x 1 ea
1.8 ml x 6 ea
Proteinase K powder, lyophilized
5 mg x 1 ea
25 mg x 2 ea
RNase A powder, lyophilized
6 mg x 1 ea
24 mg x 2 ea
*Mini - 8rxn, Midi - 1rxn **Mini –100 rxn, Midi –15 rxn, Maxi –8 rxn

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III. Storage
MagListo
TM 5M Genomic DNA Extraction Kit should be stored dry at room temperature. It can be stored for
up to 2 years if it remains sealed. MagListo
TM 5M Genomic DNA Extraction Kit provides optimized Buffer ②
(Binding) and Buffer ③(1st Washing) which is poisonous and hazardous. Please, wear gloves and goggle
eye protection when working with Buffer ②(Binding) and Buffer ③(1st Washing).
IV. 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.
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, tips
and materials should be processed and disposed of according to applicable and appropriate regulations
of the municipality/government in which this product is being used. A user must also be equipped with
basic experimental techniques required for correct execution of the experiments described in this 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 the unlicensed use of a patented application.
VI. Warranty and Liability
All BIONEER products are manufactured and tested under strict quality control protocols. BIONEER
guarantees the quality of all directly manufactured products during the warranty period of one (1) year from
the date of purchase. If any issues are discovered relating to compromise in product quality, immediately
contact BIONEER’s Customer Service Center (sa[email protected]).

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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 to BIONEER in written form within 30
days of occurrence.
VII. Technical Assistance
At Bioneer, we pride ourselves on being responsive to your needs. If you have any questions or would like
to find out more information about MagListo ™ products, please contact us. We look forward to hearing
from you!
Technical Support
For all technical questions and troubleshooting on Bioneer products and applications.
Tel: [phone]
Email: [e-mail]
In North America
Tel: [phone]
Email:[e-mail]
VIII. Quality Management
Every aspect of our quality management system from product development, production to quality
assurance and supplier qualification meets the world-class standards. Each lot of MagListo ™ 5M Genomic
DNA Extraction Kit is carefully tested by the quality control team.

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IX. Product Specifications
Sample Type
Mini
(Typical yield)
Midi
(Typical yield)
Maxi
(Typical yield)
Whole blood
200 μl ( < 10 μg)
2 ml ( < 80 μg)
4 ml ( < 150 μg)
Cultured cell
~ 1 x 106 ( < 12 μg)
~ 5 x 106 (< 60 μg)
~ 1 x 107 (< 120 μg)
Animal tissue
~ 25 mg ( < 10 μg)
~ 100 mg ( < 40 μg)
~ 250 mg (< 120 μg)
Bacteria
(Gram (-), (+))
~ 1 x 109 (< 15 μg)
~ 5 x 109 (< 80 μg)
~ 1 x 1010 (< 150 μg)
Expected purity
A260/280 > 1.8, A260/230 > 1.4
* The yield of low copy cell could be less than the figures shown in the table.
** For cultured cells, lower cell numbers can be used to a minimal cell number of ~104for “micro” scale
extraction.
Small amount of sample
MagListo
TM 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 (< 1x104) 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
starting material. Ensure that the carrier DNA does not interfere with your downstream application. Carrier
RNA can be removed later by RNase digestion. Refer to “DNA Extraction from Cultured Cell for Micro”
which is optimized protocol for extraction of DNA from low copy cell in page 10
X. Principle
MagListo
TM 5M Genomic DNA Extraction Kit is designed for the extraction of genomic DNA from a variety
of sources including high molecular weight up to 40 Kb (Note: this is often for most DNA based
application). The overall principle is based on adsorption of DNA onto the magnetic nano bead by
chaotropic salt. For instance, chaotropic agents in Buffer ②(Binding) contains guanidine hydrochloride,
as which removes water molecules around DNA and silica coated magnetic bead surface resulting in
genomic DNA then being captured by magnetic beads. The magnetic nano bead and nucleic acid
complexes are pulled and fixed on the tube wall using a magnetic force, followed by washing with ethanol

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to remove debris and excessive salts. The captured nucleic acids are then eluted by Buffer ⑤(Elution), an
aqueous solution with optimal pH.
Sample Lysis Binding Washing Elution
XI. 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
7. MagListo
TM Magnetic Separation Rack
Magnetic Separation Rack Choice
Tube
MagListo
TM Magnetic Separation Rack
1 ml tube with 8-cap strip
MagListo
TM-8Ch Magnetic Separation Rack
(Cat. no. TM-1000)
1.5 ml or 2 ml microcentrifuge tube
MagListo
TM-2 Magnetic Separation Rack
(Cat. no. TM-1010)
15 ml tube
MagListo
TM-15 Magnetic Separation Rack
(Cat. no. TM-1020)
50 ml tube
MagListo
TM-50 Magnetic Separation Rack
(Cat. no. TM-1030)
(Note) Please refer to the ordering information table on the latter part of the manual which
contains the appropriate catalog number for specific size of tubes.

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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℃.
3. Buffer ②(Binding) contains chaotropic salt. You should take the appropriate laboratory safety
precautions and wear gloves when handling.
4. Add the correct amount of absolute ethanol to Buffer ③(1st Washing) and Buffer ④(2nd Washing).
a. DNA Extraction from Whole Blood for Mini/Midi/Maxi scale
1. Add 20 μl (mini)/ 100 μl (midi)/ 200 μl (maxi) of proteinase K (see “Before you begin”) to each specific
tube format.
a. (Mini) Add proteinase K to a 1.5 ml or 2 ml tube.
b. (Midi) Add proteinase K to a 15 ml tube.
c. (Maxi) Add proteinase K to a 50 ml tube.
2. Apply 200 μl (mini)/ 2 ml (midi)/ 4 ml (maxi) of whole blood and buffy coat to the tube containing
proteinase K.
(Note) If the sample volume is less than indicated volume above, make the total volume 200 μl (mini)/
2 ml (midi)/ 4 ml (maxi) by adding 1X PBS to achieve maximum lysis efficiency and yield.
3. (Lysis: 3-4) Add 200 μl (mini)/ 2 ml (midi)/ 4 ml (maxi) 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.
4. Incubate at 60℃for 10 min.
5. (DNA Precipitation) Add 400 μl (mini)/ 4 ml (midi)/ 8 ml (maxi) of absolute ethanol and mix well using a

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vortex mixer or pipetting.
6. (DNA binding with Magnetic Nano Bead: 6-8) Add 100 μl (mini)/500 μl (midi)/ 1 ml (maxi) of Magnetic
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
TM
- 2 (mini)/ MagListo
TM
- 15 (midi)/ MagListo
TM
- 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.
8. Without removing the tube from MagListo
TM, carefully pour the supernatant out and completely remove
the remaining supernatant using a paper towel. In process, the magnetic crude pellet remains attached
to the side of tube.
-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.
9. (1st washing: 9-12) Detach the magnet plate from MagListo
TM rack. Add 500 μl (mini)/ 3 ml (midi)/ 5
ml (maxi) of Buffer ③(1st Washing) to the tube. Close the cap and mix by vortex mixer until the beads
are fully resuspended.

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-Detachment
Push up the magnet plate gently.
10. Attach the magnet plate to MagListo
TM rack and invert the tube 3 ~ 4 times gently until the beads
tightly bind to the magnet.
11. Without removing the tube from MagListo
TM rack, pour the supernatant out and remove the remaining
supernatant using a paper towel by blotting.
12. Repeat the above step 9 ~ 11 by adding 500 μl (mini)/ 3 ml (midi)/ 6 ml (maxi) of Buffer ③(1st
Washing) for additional washing to remove pigment and other debris.
13. (2nd washing) Repeat the above step 9 ~ 11 by adding 700 μl (mini)/ 5 ml (midi)/ 10 ml (maxi) of
Buffer ④(2nd Washing) for additional washing.
14. (3rd washing) Repeat the above step 9 ~ 11 by adding 700 μl (mini)/ 5 ml (midi)/ 10 ml (maxi) of
absolute ethanol for additional washing.
15. (Drying) Completely dry the beads with the tube open and use a heat gun or a blow dryer for 1 min
(mini)/ 3 min (midi)/5 min (maxi) 3 cm away from the top of the tube.
(Note) Without using a heat gun or a blow dryer, place the rack lying down in a dry oven at 60℃for 10
min (mini)/ 20 min (midi)/30 min (maxi).
16. (Elution: 16-20) Add 100 μl (mini)/ 500 μl (midi)/ 1 ml (maxi) of Buffer ⑤(Elution) or distilled water to
the tube with the magnet plate detached and resuspend completely by pipetting or vortex mixer for 15
sec.
17. Incubate the tube at 60℃for 1 min.

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18. Attach the magnet plate to MagListo
TM rack and invert the tube 3 ~ 4 times gently until the beads
tightly bind to the magnet.
19. Without removing the tube from MagListo
TM rack, carefully take the supernatant containing DNA to a
sterile microcentrifuge tube.
20. Discard the used magnetic nano beads. Do not reuse the beads.
Summary of Reagents Volume in Each Step of DNA Extraction from Whole Blood
Step
Buffer
Mini
Midi
Maxi
Page
Blood (+ PBS)
200 μl
2 ml
4 ml
P. 6
Lysis
Buffer ②(Binding)
200 μl
2 ml
4 ml
P. 6
Precipitation
Absolute Ethanol
400 μl
4 ml
8 ml
P. 6
Bead Binding
Magnetic Nano Bead - DNA
100 μl
500 μl
1 ml
P. 6
1st Washing
(Repeat)
Buffer ③(1st Washing)
500 μl
3 ml
6 ml
P. 7
2nd Washing
Buffer ④(2nd Washing)
700 μl
5 ml
10 ml
P. 8
3rd Washing
Absolute Ethanol
700 μl
5 ml
10 ml
P. 8
Elution
Buffer ⑤(Elution)
100 μl
500 μl
1 ml
P. 8

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b. DNA Extraction from Cultured Cell for Micro/Mini/Midi/Maxi
1. Centrifuge the cultured cells (~ 1x104(micro)/ ~ 1x106(mini)/ ~ 5x106(midi)/ ~ 1x107 (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 1x104cells. Thus, given volume amount of this kit component is adjusted and mentioned on this
procedure.
2. Resuspend the pellet in 100 μl (micro)/ 200 μl (mini)/ 1 ml (midi, maxi) of 1X PBS and transfer them to
each specific tube format.
a. (Micro/Mini) Transfer the resuspended pellet to a 1.5 ml or 2 ml tube.
b. (Midi/Maxi) Transfer the resuspended pellet to a 15 ml tube.
3. Add 10 μl (micro)/ 20 μl (mini)/ 100 μl (midi)/ 200 μl (maxi) of proteinase K (see “Before you begin”) to
the tube.
4. If RNA-free genomic DNA is required, add up to 2 μl (micro)/ 10 μl (mini)/ 75 μl (midi), 150 μl (maxi) of
RNase A (see “Before you begin”) and incubate for 5 min at room temperature.
5. (Lysis: 5-6) Add 100 μl (micro)/ 200 μl (mini)/ 1 ml (midi, maxi) of Buffer ②(Binding) to the sample
and mix immediately and thoroughly using a vortex mixer. You must completely resuspend the sample
to achieve maximum lysis efficiency.
6. Incubate at 60℃for 10min.
7. (DNA precipitation) Add 200 μl (micro)/ 400 μl (mini)/ 2 ml (midi, maxi) of absolute ethanol and mix
well by vortex mixer or pipetting.
8. (DNA binding with Magnetic Nano Bead: 8-10) Add 100 μl (micro, mini)/500 μl (midi)/ 1 ml (maxi) of
Magnetic 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.

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9. Place the tube in MagListo
TM
- 2 (micro, mini)/ MagListo
TM
- 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.
10. Without removing the tube from MagListo
TM rack, carefully pour the supernatant out and completely
remove the remaining supernatant using a paper towel by blotting.
-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.
11. (1st washing: 11-13) Detach the magnet plate from MagListo
TM rack. Add 700 μl (micro, mini)/ 5 ml
(midi)/ 10 ml (maxi) of Buffer ③(1st Washing) to the tube. Close the cap and mix by vortex mixer until
the beads are fully resuspended.
-Detachment
Push up the magnet plate gently.
12. Attach the magnet plate to MagListo
TM stand and invert the tube 3~4 times gently until the beads

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tightly bind to the magnet.
13. Without removing the tube from MagListo
TM rack, pour the supernatant out and remove the remaining
supernatant using a paper towel by blotting.
14. (2nd washing) Repeat the above step 11 ~ 13 by adding 700 μl (micro, mini)/ 5 ml (midi)/ 10 ml (maxi)
Buffer ④(2nd Washing) for additional washing.
15. (3rd washing) Repeat the above step 11 ~ 13 by adding 700 μl (micro, mini)/ 5 ml (midi)/ 10 ml (maxi)
absolute ethanol for additional washing.
16. (Drying) Completely dry the beads with the tube open and use a heat gun or a blow dryer for 1 min
(micro, mini)/ 3 min (midi)/ 5 min (maxi) 3 cm away from the top of the tube.
(Note) Without using a heat gun or a blow dryer, place the rack lying down in a dry oven at 60℃for 10
min (micro, mini)/ 20 min (midi)/ 30 min (maxi).
17. (Elution: 17-21) Add 50 μl (micro)/ 100 μl (mini)/ 500 μl (midi)/ 1 ml (maxi) of Buffer ⑤(Elution) or
distilled water to the tube with the magnet plate detached and resuspend completely by pipetting or
vortex mixer for 15 sec.
18. Incubate the tube at 60℃for 1 min.
19. Attach the magnet plate to MagListo
TM rack and invert the tube 3~4 times gently until the beads tightly
bind to the magnet.
20. Without removing the tube from MagListo
TM rack, carefully take the supernatant containing DNA to a
sterile microcentrifuge tube.
21. Discard the used magnetic nano beads. Do not reuse the beads.

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Summary of Reagents Volume in Each Step of DNA Extraction from Cultured Cell
Step
Buffer
Micro
Mini
Midi
Maxi
Page
Cultured Cell
~ 1 x 104
~ 1 x 106
~ 5 x 106
~1 x 107
P. 10
Lysis
Buffer ②(Binding)
100 μl
200 μl
1 ml
1 ml
P. 10
DNA Precipitation
Absolute Ethanol
200 μl
400 μl
2 ml
2 ml
P. 10
Bead Binding
Magnetic Nano Bead -
DNA
100 μl
100 μl
500 μl
1 ml
P. 10
1st Washing
Buffer ③(1st Washing)
700 μl
700 μl
5 ml
10 ml
P. 11
2nd Washing
Buffer ④(2nd Washing)
700 μl
700 μl
5 ml
10 ml
P. 12
3rd Washing
Absolute Ethanol
700 μl
700 μl
5 ml
10 ml
P. 12
Elution
Buffer ⑤(Elution)
50 μl
100 μl
500 μl
1 ml
P. 12

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c. DNA Extraction from Animal Tissue for Mini/Midi/Maxi
1. (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. (Midi) Place the homogenized tissue to a 15 ml tube.
c. (Maxi) Place the homogenized tissue to a 50 ml tube.
(Note) Immediately place the weighted, fresh or frozen tissue in liquid nitrogen and grind to a fine
powder with mortar and pestle under liquid nitrogen. Final yield of DNA depends on the amount
and the type of the used tissue.
2. (Lysis: 2-6) Add 180 μl (mini)/ 1.8 ml (midi)/ 3.6 ml (maxi) of Buffer ⓛ(Lysis).
3. Add 20 μl (mini)/ 100 μl (midi)/ 200 μl (maxi) of proteinase K (see “Before you begin”) to the tube and
mix thoroughly using a vortex mixer.
4. If RNA-free genomic DNA is required, add up to 10 μl (mini)/ 75 μl (midi)/ 150 μl (maxi) of RNase A
(see “Before you begin”) and incubate for 2 min at room temperature.
5. Incubate at 60℃until the tissue is completely lysed.
6. Add 200 μl (mini)/ 2 ml (midi)/ 4 ml (maxi) of Buffer ②(Binding) to the tube and mix immediately and
thoroughly using a vortex mixer. You must completely resuspend the sample to achieve maximum lysis
efficiency.
7. (DNA precipitation) Add 400 μl (mini)/ 4 ml (midi)/ 8 ml (maxi) of absolute ethanol and mix well using a
vortex mixer or pipetting.
8. (DNA binding with Magnetic Nano Bead: 8-10) Add 100 μl (mini)/500 μl (midi)/ 1 ml (maxi) of
Magnetic 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.

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9. Place the tube in MagListo
TM
- 2 (mini)/ MagListo
TM
- 15 (midi)/ MagListo
TM
- 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.
10. Without removing the tube from MagListo
TM rack, carefully pour the supernatant out and completely
remove the remaining supernatant using a paper towel by blotting.
-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.
11. (1st washing: 11-13) Detach the magnet plate from MagListo
TM rack. Add 700 μl (mini)/ 5 ml (midi)/ 10
ml (maxi) of Buffer ③(1st Washing) to the tube. Close the cap and mix by vortex mixer until the beads
are fully resuspended.
-Detachment
Push up the magnet plate gently.

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12. Attach the magnet plate to MagListo
TM stand and invert the tube 3 ~ 4 times gently until the beads
tightly bind to the magnet.
13. Without removing the tube from MagListo
TM rack, pour the supernatant out and remove the remaining
supernatant using a paper towel by blotting.
14. (2nd washing) Repeat the above step 11 ~ 13 by adding 700 μl (mini)/ 5 ml (midi)/ 10 ml (maxi) Buffer
④(2nd Washing) for additional washing.
15. (3rd washing) Repeat the above step 11 ~ 13 by adding 700 μl (mini)/ 5 ml (midi)/ 10 ml (maxi)
absolute ethanol for additional washing.
16. (Drying) Completely dry the beads with the tube open and use a heat gun or a blow dryer for 1 min
(mini)/ 3 min (midi)/5 min (maxi) 3 cm away from the top of the tube.
(Note) Without using a heat gun or a blow dryer, place the rack lying down in a dry oven at 60℃for 10
min (mini)/ 20 min (midi)/30 min (maxi).
17. (Elution: 17-21) Add 100 μl (mini)/ 500 μl (midi)/ 1 ml (maxi) of Buffer ⑤(Elution) or distilled water to
the tube with the magnet plate detached and resuspend by pipetting or vortex mixer for 15 sec.
18. Incubate the tube at 60℃for 1 min.
19. Attach the magnet plate to MagListo
TM rack and invert the tube 3~4 times gently until the beads tightly
bind to the magnet.
20. Without removing the tube from MagListo
TM rack, carefully take the supernatant containing DNA to a
sterile microcentrifuge tube.
21. Discard the used magnetic nano beads. Do not reuse the beads.

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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
Lysis
Buffer ⓛ(Lysis)
180 μl
1.8 ml
3.6 ml
P. 14
Buffer ②(Binding)
200 μl
2 ml
4 ml
P. 14
DNA
Precipitation
Absolute Ethanol
400 μl
4 ml
8 ml
P. 14
Bead Binding
Magnetic Nano Bead –DNA
100 μl
500 μl
1 ml
P. 14
1st Washing
Buffer ③(1st Washing)
700 μl
5 ml
10 ml
P. 15
2nd Washing
Buffer ④(2nd Washing)
700 μl
5 ml
10 ml
P. 16
3rd Washing
Absolute Ethanol
700 μl
5 ml
10 ml
P. 16
Elution
Buffer ⑤(Elution)
100 μl
500 μl
1 ml
P. 16