www.toyobo.co.jp/e/bio

F0981K

Instruction manual MagExtractor-Genome-2004

MagExtractor -GenomeNPK-101

100 preparations

Store at 4°C

Contents[1]

Introduction[2]

Components[3]

Materials required[4]

Protocol1. Purification from whole blood or cultured cells
2. Purification from animal tissue or mouse tail[5]

Troubleshooting[6]

References[7]

Related products

CAUTIONAll reagents in this kit are intended for research purposes. Do not use for diagnostic or clinical purposes. Please observe
general laboratory precaution and utilize safety while using this kit.
JAPANTOYOBO CO., LTD.
Tel(81)-6-6348-3888
www.toyobo.co.jp/e/bio
tech_osaka@toyobo.jp

CHINA
TOYOBO (SHANGHAI BIOTECH, CO., LTD.)
Tel(86)-21-58794900

FOR RESEARCH USE ONLY.

NOT FOR HUMAN OR DIAGNOSTIC USE.

1

www.toyobo.co.jp/e/bio[ 1 ] Introduction

Description
MagExtractor -Genome- provides a simple and reliable method for the rapid purificationof genomic DNA from various specimens (e.g. whole blood, cultured cells or animal tissuesetc) using magnetic silica beads. This kit is based on binding properties of DNA to a silicasurface in the presence of chaotropic agents 1)2). The purified genomic DNA can be useddirectly for PCR experiments.

Lysis

Binding

Washing

ElutionFig. 1 Principle of purification

Features-This kit can be applied to extraction of DNA from whole blood specimens. A leukocyte orlymphocyte separation step is not necessary.
-This kit does not contain hazardous substances, such as phenol or chloroform.
-Purified genomic DNA can be eluted in sterilized water. Therefore, the purified DNA
samples can be applied directly to other methods, such as PCR, etc.
-This kit is suitable for high-throughput extraction of genomic DNA from various
specimens. The following table shows the typical yield and purity in each case.
SampleYieldPurity(A260/280)

Application

Blood
2 µg/100μL whole blood
1.8 ± 0.1
PCR
Cultured cells 3 µg/5x105 HeLa cells
Tissue5 µg/5 mg tissue (swine testis)
Mouse tail3 µg/2 mm tailNotes
In the case of tissues or mouse tails, homogenization and/or lysis steps are necessaryprior to extractionn.

[ 2 ] Components

This kit contains the following components for 100 preparations. All reagents should be
stored at 4 °C.
Lysis & Binding Solution
Washing Solution
Magnetic Beads

100 mL
200 mL
5 mL

Caution:- The “Lysis & Binding Solution” and “Washing Solution” contain chaotropic salt, which
is an irritant. Take appropriate laboratory safety measures and wear gloves when handling.
If contact with skin occurs, wash thoroughly with water. If the eyes get affected, flush
thoroughly for 15 min. with cool water, and consult a physician.
Notes:-All reagents should be used at room temperature for extraction.
-If used within one month or less, all components can be stored at room temperature(≤25°C).
JAPANTOYOBO CO., LTD.
Tel(81)-6-6348-3888
www.toyobo.co.jp/e/bio
tech_osaka@toyobo.jp

CHINA
TOYOBO (SHANGHAI) BIOTECH, CO., LTD.
Tel (+86)-21-58794900

FOR RESEARCH USE ONLY.

NOT FOR HUMAN OR DIAGNOSTIC USE.

1

www.toyobo.co.jp/e/bio[3]

Materials required

The following materials are required for purification.
(1) Reagents-Sterilized water-70% Ethanol(2) Instruments-Tube mixer-Magnetic stand[4]ProtocolFig.2
Magnetic standMagical Trapper (Code No.MGS-101)

1. Purification from whole blood or cultured cells<Standard protocol>(1) Preparation of specimens-Whole blood
Dispense 100 μL of whole blood into 1.5-mL microtubes.
(Whole blood can be used directly.)-Cultured cells
Collect cultured cells by centrifugation in PBS(-), and dispense 1x105-106 cells/100
μL PBS(-) into 1.5-mL microtubes.
(2) Add 750 μL Lysis & Binding Solution.
(3) [Binding] Add 40 μL Magnetic Beads and mix well for 10 minutes using a tubemixer.
NoteSuspend Magnetic Beads completely prior to use.
(4) Place each tube into the magnetic stand. The magnet
will attract the magnetic beads, separating from thespecimen solution.
(5) After magnetic capture, carefully remove thesupernatant.

Fig. 3Magnetic separation(6) [Washing] Add 900 μL Washing Solution to the beads and vortex for 5 seconds(Maximum speed).
(7) Place the tube on a magnetic stand and collect the beads for 30 seconds.
(8) After magnetic capture, carefully remove the supernatant.
(9) [Washing] Repeat (6) - (8)(10) [Washing] Add 900 μL 70% Ethanol and vortex 5 seconds (Maximum speed).

JAPANTOYOBO CO., LTD.
Tel(81)-6-6348-3888
www.toyobo.co.jp/e/bio
tech_osaka@toyobo.jp

CHINA
TOYOBO (SHANGHAI) BIOTECH, CO., LTD.
Tel (+86)-21-58794900

FOR RESEARCH USE ONLY.

NOT FOR HUMAN OR DIAGNOSTIC USE.

2

www.toyobo.co.jp/e/bio(11) Place the tube on a magnetic stand and collect the beads for 30 seconds with themagnet.
(12) After magnetic capture, carefully remove the supernatant.
(13) [Washing] Repeat (10) - (12)(14) [Elution] Add 100 μL sterilized water and mix well for 10 minutes using a tube
mixer.
(15) Place the tube in the magnetic stand.
(16) Collect the supernatant and place in a fresh tube.

2. Purification from animal tissue or mouse tails(1) Pretreatment of specimens
Animal tissue or mouse tails should be treated with either of the following methods.

(A) Homogenization of specimens in Lysis & Binding Solution.
Tissue (≤ 10mg) or mouse tail (2-5 mm)

 850 μL Lysis & Binding Solution
Homogenization

Centrifuge at 10,000 rpm for 5 minutes

Supernatant 850 μL (Pretreated solution)(B) Homogenization after Proteinase K treatment
Tissue (≤ 10mg) or mouse tail (2-5 mm)

 90 μL Proteinase K buffer [100 mM NaCl, 10 mM Tris-HCl (pH 8.0),25mM EDTA]

 5 μL 10 mg/mL Proteinase K (30 U/mg)
 5 μL 10% SDS
Incubate at 55°C for 6-18 hr. (Mix 2-3 times halfway through incubation)

Centrifuge at 10,000 rpm for 5 minutes

Supernatant 100 μL
  750 μL Lysis & Binding Solution
Pretreated solution 850 μL(2) Perform the purification steps (3)-(16) in Standard protocol.
JAPANTOYOBO CO., LTD.
Tel(81)-6-6348-3888
www.toyobo.co.jp/e/bio
tech_osaka@toyobo.jp

CHINA
TOYOBO (SHANGHAI) BIOTECH, CO., LTD.
Tel (+86)-21-58794900

FOR RESEARCH USE ONLY.

NOT FOR HUMAN OR DIAGNOSTIC USE.

3

www.toyobo.co.jp/e/bio[ 5 ] Troubleshooting
Symptom

CauseSolution

Excess specimen
Insufficient homogenization

Low yield

Poor amplification on
PCR[ 6 ] References

Ethanol inhibition

Excess samples decrease DNA yields. Useappropriate amount of specimen.
-Insufficient homogenization of tissue or mouse
tails decreases DNA yields. Prolong thehomogenization step.
-In the case of formalin-fixed tissue, Proteinase K
pretreatments might be effective.
-In the case of gram-negative bacteria or yeast,pretreatments using specific enzymes (e.g.,zymolyase for yeast) are necessary. Gramnegative bacteria (e.g., E. coli) can be useddirectly.
Decrease the amount of DNA solution for PCR byup to 20% of reaction volume. Purified DNA
samples contain residual amounts of ethanol fromthe washing steps.
The ethanol can be evaporated by heating sample
at 75°C for 5 minutes.

1) B. Vogelstein and D. Gillespie, Proc. Natl. Acad. Sci. USA. 76: 615-619 (1979)
2) R. Boom, C. J. A. Sol, M. M. M.Salimans, C. L. Wertheim-van Dillen, P. M. E. Dillen
and J. van der Noordaa, J. Clin. Microbiol., 28: 495-503 (1990)[ 7 ] Related products
Product name

Package1 piece

Magnetic standCode No.
MGS-101

Magical Trapper

JAPAN
TOYOBO CO., LTD.
Tel(81)-6-6348-3888
www.toyobo.co.jp/e/bio
tech_osaka@toyobo.jp

CHINA
TOYOBO (SHANGHAI) BIOTECH, CO., LTD.
Tel (+86)-21-58794900

FOR RESEARCH USE ONLY.

NOT FOR HUMAN OR DIAGNOSTIC USE.

4