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F0985K

Instruction manual MagExtractor -Plant Genome- 2004

MagExtractor -Plant GenomeNPK-501

100 preparations

Store at 4 °C

Contents[1]

Introduction[2]

Components[3]

Materials required[4]

Protocol1. Pretreatment of plant specimens
2. Purification[5]

Troubleshooting[6]

References[7]

Related products

CAUTIONAll reagents in this kit are intended for research purposes. Do not use for diagnosis or clinical purposes. Please observe generallaboratory precautions, and follow safety guidelines 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.

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

Introduction

Description
MagExtractor -Plant Genome- provides a simple and reliable method for the rapid
purification of genomic DNA from various plant specimens, e.g. leaf and cultured cells,
etc, using magnetic silica beads. This kit is based on binding properties of DNA to a silica
surface in the presence of chaotropic agents1). Purified genomic DNA can be used directlyfor PCR experiments.

Fig. 1. Principles of the purification process.

Features- Purified genomic DNA can be used directly for PCR.
- The pretreatment step of this kit is effective in removing polysaccharides.
- This kit is suitable for the high-throughput extraction of genomic DNA from various plant
specimens. The following tables show typical sample amounts, yields, and applications
for purified genomic DNA.
Table 1. Typical sample amounts and applications
SampleSample amount
Plant specimens
0.01 - 0.1g
(leaf, cultured cells, etc)

Application
PCR

Table 2. Typical yields from various types of plant tissues (0.1 g).
SampleYield (µg)
Tabacco (leaf)
3-5
Tabacco (cultured cells)
2-3
Rice (leaf)
3-5
Arabidopsis (leaf)
1
Tomato (leaf)
3
Pumpkin (leaf)
3
Maize (leaf)
3-4
Notes
The yield depends on the types of species, tissues, and sample conditions that are used.
Table 2 shows typical yields from various plant tissues.

JAPANTOYOBO CO., LTD.
Tel(81)-6-6348-3888
www.toyobo.co.jp/e/bio
tech_osaka@toyobo.jpCHINATOYOBO (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[ 2 ] Components
This kit includes the following components for 100 preparations. All reagents, except for
the lysis solution should be stored at 4 °C. The lysis solution should be stored at room
temperature (20 - 30 °C) once dissolved.
Lysis Solution
Binding Solution
Washing Solution
Magnetic Beads

40 mL
90 mL
200 mL
6 mL

Caution:- The “Binding Solution” and “Washing Solution” contain chaotropic salts, which areirritant. Follow appropriate laboratory safety guidelines, and wear gloves when handlingthe reagents. 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.
- The “lysis solution” generates precipitates consisting of detergents at low temperature. If
precipitates are formed, dissolve the precipitates according to the following method. Be
cautious as this solution tends to generate transparent precipitates.
1. Incubate the bottle in a hot water bath at 55 °C for 10 min.
2. Mix the contents of the bottle gently by inverting.
3. Incubate the bottle in a hot water bath at 55 °C for 5 min.
4. Mix the contents of the bottle gently by inverting until the precipitates disappear.
Notes- All reagents at room temperature should be used for extraction.
- For short-term storage (≤ 1 month), all components can be kept at room temperature.

[3]

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

The following materials are required for purification.
1.

Reagents- Chloroform / Isoamylalcohol (24:1)- TE Buffer (10 mM Tris-HCl, pH 8.0, 1 mM EDTA)- 70% ethanol- Sterile water- Liquid nitrogen

2.

Instruments- Tube mixer- Magnetic stand- Microcentrifuge- Heating block (65 °C)

Fig.2
Magnetic standMagical Trapper (Code No.MGS-101)

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

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www.toyobo.co.jp/e/bio[4]Protocol

1. Pretreatment of plant specimens
The following pretreatment steps must be performed prior to purification.
(1) Disrupt plant specimens thoroughly using liquid nitrogen.
Notes
Plant specimens should be disrupted thoroughly until the specimens are powdery.
(2) Transfer the specimen (0.01 - 0.1 g) into a chilled 1.5-mL microtube using a chilledspatula.
(3) Add 300 μL lysis solution.
Notes
If precipitates have formed in the lysis solution, dissolve the precipitates according
to the method described in [2].
(4) Mix thoroughly using a vortex mixer for 10 sec.
(5) Incubate at 65 °C for 10 min. (vortex the tube every 3 - 4 min.).
(6) Add 300 μL chloroform/isoamylalcohol (24:1) to the tube.
(7) Mix thoroughly by inverting the tube 3 - 5 times.
(8) Centrifuge at 12,000 rpm for 1 min.
Notes
Incomplete separation may be improved by an extra 2 - 3 min. centrifugation.
(8) Carefully transfer the upper solution (250 μL) to a fresh 1.5-mL microtube.
(9) Add 600 μL Binding Solution.
Notes
If the volume of the upper solution is less than 100 μL, add 750 μL BindingSolution.

2. Purification(1) Binding. Add 40 μL Magnetic Beads to the pretreated solution (see abovesection) and mix thoroughly for 1 min. using a vortex mixer.
Notes
Suspend the magnetic beads completely prior to use.
(2) Place each tube on a magnetic stand. The magnet
will attract the magnetic beads, separating themfrom the specimen solution.
(3) After magnetic capture, carefully remove thesupernatant.

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Fig. 3.
Magnetic separation

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www.toyobo.co.jp/e/bio(4) Washing. Add 900 μL Washing Solution to the beads, and mix thoroughly for
5 sec. using the vortex mixture.
(5) Place each tube on a magnetic stand, and collect the beads with the magnet.
(6) After magnetic capture, carefully remove the supernatant.
(7) Washing. Repeat (4) - (6)(8) Washing. Add 900 μL 70% ethanol, and mix thoroughly for 5 sec using thevortex mixture.
(9) Place each tube on a magnetic stand, and collect the beads with the magnet.
(10) After magnetic capture, carefully remove the supernatant.
(11) Washing. Repeat (8) - (10)(12) Elution. Add 100 μL TE Buffer and mix thoroughly for 1 min.
(13) Place the tube on a magnetic stand.
(14) Collect the supernatant into a fresh tube.
Notes
Purified DNA samples contain small amounts of ethanol from the washing steps.
Ethanol can be evaporated by heating at 75 °C for 5 min, and leaving the lid of
the microtube opened.

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.

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

Cause

Solution

Excess specimen
Insufficient disruption
Low yield

Poor amplification at
PCR

Incomplete separation at thecentrifugation step

Inhibition by ethanol

Excess samples decrease DNA yields. Useappropriate amounts of plant tissues.
Plant specimens should be disrupted thoroughlyuntil the specimens are powdery. Be cautious thatthe specimens do not thaw.
-Thoroughly reconstitute disrupted specimens andlysis solution. Prolong the vortexing time from10 s to 1 min.
- Increase the amount of lysis solution up to 350 450 μL .
Decrease the amount of DNA solution for PCR
using a decrement up to 20% of the reaction
volume. Purified DNA samples contain small
amounts of ethanol from the washing steps.
Ethanol can be evaporated by heating at 75 °C for
5 min, and leaving the lid of the microtube
opened.

1) B. Vogelstein and D. Gillespie, Proc. Natl. Acad. Sci. USA. 76: 615-619 (1979)[ 6 ] Reference[ 7 ] Related products

Product namePackage
1 piece

Magnetic standa

Code 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.

5