Version：01/2021

Magbead Blood Spots DNA Kit

Catalog Number：CW2504S（96 preps）

Storage Condition：Room Temperature（15-30℃）
Kit Components
CW2504S
Component
96 preps
Buffer WL 36 mL
Buffer KL 50 mL
Buffer GW1（concentrate） 80 mL
Buffer GW2（concentrate） 50 mL
Buffer EB 30 mL
Proteinase K 2×25 mg
Proteinase K Storage Buffer 2×1.25 mL
Magbeads PN 2×1 mL

Introduction
This product is for scientific research only, not for clinical diagnosis and other purposes.
The kit provides a simple, rapid and efficient method for DNA extraction from blood slices,
suitable for extraction from blood slices
Take genomic DNA. In the presence of high salt, DNA binds to the surface of silicon-based
coated Magbeads. After rinsing, the highly purified DNA is eluted in Buffer EB or deionized
water. The purified DNA has good purity (A260/280 ratio between 1.7-1.9) and high integrity
(>15 KB), which can be used for downstream experiments such as second-generation
sequencing, quantitative PCR and chip detection.
The kit can be matched with CWE2100 32-channel nucleic acid extractor and CWE9600 96-
channel nucleic acid extractor
With the use of simple, fast high-throughput extraction, greatly reduce the experimenter's
workload and experimental error.
Instruments and reagents provided by user

1）Constant temperature blender——Cat.No.：CW2593

2）2/15 mlMagnetic Stand——Cat.No.：CW2594
3）32 channels Nucleic Acid Extractor ——Cat.No.：CWE2100
4）96 channels Nucleic Acid Extractor ——Cat.No.：CWE9600
5）96 DW Plate——Cat.No.：CW2523
6）8 channel Comb——Cat.No.：CW2524
7）Spin tips pack——Cat.No.：CW2532
8）ethanol

Preparation and important points for attention before experiment

1. 1.25 ml Proteinase K Storage Buffer was added to 25 mg Proteinase K to

dissolve it, and then stored at -20℃. The Proteinase K solution should not be
placed at room temperature for a long time to avoid repeated freezing and thawing,
so as not to affect its activity.
2. Before the first use, anhydrous ethanol was added to Buffer GW1 and Buffer
GW2 according to the reagent bottle label and marked.
3. Magbeads are strictly prohibited from freezing and centrifugation. Freezing and
centrifugation may cause irreversible damage to Magbeads.

Procedure
Ⅰ、Manual single-tube operation

1. Take one blood plaque with a diameter of 6 mm or four blood plaques with a
diameter of 3 mm (according to the actual situation) from the blood plaque and put
them into a 2.0 mL centrifuge tube.
2. Add 20 μ L Proteinase K and 300 μ L Buffer WL to the centrifuge tube, then put
the centrifuge tube in the constant temperature mixer at 56℃ and 1200 RPM for
shock lysis for 30 minutes to form Lysate, remove the centrifuge tube from the
constant temperature mixer, centrifuge briefly, and take the supernatant.
Note: If no constant temperature mixer is available, place the centrifugal tube
in a 65℃ water bath for incubation for 20 minutes after vortex oscillation for
10 seconds, and vortex oscillation for 10 seconds every 5 minutes during this
period.
3. Drain the supernatant into a new 2.0ml centrifuge tube, and add 450 μ L Buffer
KL and 20 μ L Magbeads PN. After that, the centrifuge tube was placed on the
constant temperature blender at 25℃ and 1600 RPM for shock cracking for 10
minutes or the centrifuge tube was continuously reversed and mixed for 15
minutes.
4. Place the centrifugal tubes on the magnetic rack and stand for 1 minute. After
Magbeads are completely adsorbed on the side wall of the centrifugal tubes,
discard them completely remove solution (keep centrifuge tube fixed on magnetic
holder).
5. Remove the centrifugal tube from the magnetic rack, Add 750 μ L Buffer GW1
(check whether anhydrous ethanol has been added before use). After 1 minute of
scroll shaking or 5 seconds of scroll shaking, place the beads in a 1600 RPM
constant temperature mixer at 25 ° C and shake for 2 minutes. (Make sure the
Magbeads are in the uniform state during the shaking.) After that, put the
centrifugal tube on the magnetic frame and stand for 1 minute. After Magbeads are
completely absorbed on the side wall of the centrifugal tube, gently reverse the
magnetic frame and move away
6. Discard the solution completely after washing the impurities on the core tube
cover (keep the centrifugal tube fixed on the magnetic rack).
7. Repeat Step 5.
Remove the centrifuge tubes from the magnetic support and add 750 μ L Buffer
GW2 (check whether anhydrous ethanol has been added before use). After 1
minute of scroll shaking or 5 seconds of scroll shaking, place the beads in a 1600
RPM constant temperature mixer at 25℃ and shake for 2 minutes. (Make sure the
Magbeads are in the uniform state during the shaking.) After that, put the
centrifugal tube on the magnetic frame and stand for 1 minute. After Magbeads are
completely absorbed on the side wall of the centrifugal tube, gently reverse the
magnetic frame and move away
After washing the impurities on the core tube cover, discard the solution completely
(keep the centrifugal tube fixed on the magnetic rack).
8. Repeat Step 7.
9. Keep the centrifugal tube fixed on the magnetic rack, use a pipette to further
remove the solution on the bottom and cover of the centrifugal tube, and then leave
the tube at room temperature for 5-10 minutes to make the ethanol volatilize clean.
10. Remove the centrifugal tube from the magnetic rack and add 50-200 μ L Buffer
EB. The magnetic beads were completely suspended in the eluent by vortex
oscillation, and then the beads were placed in a constant temperature blender at
56℃ and 1600 RPM for oscillation elution for 10 minutes, or the centrifuge tube was
placed in a water bath at 56℃ for incubation for 10 minutes, during which the beads
were oscillated for 10 seconds every 3 minutes.
11. Place the centrifuge tubes on the magnetic rack and stand for 2 minutes. After
Magbeads are completely adsorbed on the side wall of the centrifuge tubes,
transfer the eluent to a new centrifuge tube with a pipette and store it at -20℃ for
later use.

Ⅱ、Matching with CWE2100

1. Take one blood plaque with a diameter of 6 mm or four blood plaques with a
diameter of 3 mm (according to the actual situation) from the blood plaque and put
them into a 2.0 mL centrifuge tube.
2. Add 20 μ L Proteinase K and 300 μ L Buffer WL into the centrifuge tube, and then
put the centrifuge tube into the constant temperature blender at 56℃ and 1200
RPM for 30 minutes for shock cracking to form Lysate;
3. Add reagents to the 96DW deep-hole plate according to the table below.

Position Reagent
Lysate: All
1&7 Colume Buffer KL: 450 μl
Magbeads PN: 20 μl
2&8 Colume Buffer GW1: 750 μl
3&9 Colume Buffer GW1: 750 μl
4&10 Colume Buffer GW2: 750 μl
5&11 Colume Buffer GW2: 750 μl
6&12 Colume Buffer EB: 100 μl

4. Place the deep-hole plate and magnetic sleeve with the reagent in the
corresponding position of CWE2100, run the blood plate extraction program,
and take out the deep-hole plate and magnetic sleeve about 30 minutes after
the program is finished.
5. The elution products in column 6&12 of the deep-hole plate were transferred
to a 1.5ml centrifuge tube for preservation at low temperature.
 Ⅲ、Matching with CWE9600

1. Take one blood plaque with a diameter of 6 mm or four blood plaques with a
diameter of 3 mm (according to the actual situation) from the blood plaque and put
them into a 2.0 mL centrifuge tube.
2. Add 20 μ L Proteinase K and 300 μ L Buffer WL into the centrifuge tube, and then
put the centrifuge tube into the constant temperature blender at 56℃ and 1200
RPM for 30 minutes for shock cracking to form Lysate;
3. Add reagents to the 96DW deep-hole plate according to the table below.
Position Reagent
Lysate: All
Plate 1 Buffer KL: 450 μl
Magbeads PN: 20 μl
Plate 2 Buffer GW1: 750 μl
Plate 3 Buffer GW1: 750 μl
Plate 4 Buffer GW2: 750 μl
Plate 5 Buffer GW2: 750 μl
Plate 6 Buffer EB: 100 μl

4. Place the deep-hole plate and magnetic sleeve with the reagent in the
corresponding position of CWE9600, run the blood film extraction program, and
take out the deep-hole plate and magnetic sleeve about 30 minutes after the
program finishes.
5. The elution products in Plate 6 were transferred to a 1.5 mL centrifuge tube
and stored at low temperature.

This product is for scientific research only, not for

clinical diagnosis and other purposes.