www.toyobo.co.jp/e/bio

F0984K

Instruction manual MagExtractor -Viral RNA- 2004

MagExtractor -Viral RNANPK-401F 100 preparations

Store at 4 °C

Contents[1]

Introduction[2]

Components[3]

Materials required[4]

Protocol[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/bio[ 1 ] Introduction

Description
MagExtractor -Viral RNA- provides a simple and reliable method for the rapid purificationof viral RNA from serum or plasma specimens using magnetic silica beads. This kit isbased on the property that RNA can be absorbed onto a silica surface in the presence ofchaotropic agents 1) 2) and an RNA-binding accelerator. The purified viral RNA can be useddirectly for RT-PCR experiments.

LysisBinding

WashingElutionFig. 1. Principles of the purification process.

Features- This kit is suitable for the high-throughput extraction of viral RNA from serum or plasma
specimens using magnetic silica beads.
- This kit does not contain hazardous substances such as phenol or chloroform.
- No ethanol is used in the washing steps.

[ 2 ] Components
This kit includes the following components for 100 preparations. All components should
be stored at 4 °C.
Lysis & Binding Solution*
Washing Solution I
Washing Solution IIElution Solution
Magnetic Beads

77 mL
154 mL
200 mL
10 mL
8 mL* Lysis & Binding Solutions and 2-Mercaptoethanol should be mixed at a ratio of 100:1
prior to use. 2-ME is not supplied with this kit.
Caution:- The “Lysis & Binding Solution” and “Washing Solution I” contain chaotropic salts, which
are irritant. Follow appropriate laboratory safety measures, and wear gloves whenhandling the 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.

[3]

Materials required

The following materials are required.
- 2-Mercaptoethanol (2-ME)- Magnetic stand- Heating block (set at 65 °C).
- Tube mixer

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

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

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

FOR RESEARCH USE ONLY.

1NOT FOR HUMAN OR DIAGNOSTIC USE.

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

Protocol1. Preparation of additional reagents (not provided)(1) Lysis & Binding solution (2-ME)
The Lysis & Binding Solution and 2-mercaptoethanol (2-ME) should be mixed at aratio of 100:1 prior to use. If precipitates form, dissolve the precipitates at roomtemperature. 2-ME is not supplied with this kit.
(2) Washing Solutions I and II are stored at room temperature, and should be used forpurification.

2.

Purification(1) Add 700 μL Lysis & Binding Solution (2-ME) to 100-300 μL serum or plasma
samples.
(2) [Binding] Add 50μL Magnetic Beads, and mix thoroughly for 10 min. using atube mixer.
Notes
Suspend the magnetic beads completely prior to use.
(3) Place each tube on a magnetic stand. The magnet
will attract the magnetic beads, separating themfrom the specimen solution.
(4) After magnetic capture, carefully remove thesupernatant.

Fig. 3Magnetic separation(5) [Washing] Add 700 μL Washing Solution I to the beads, and mix thoroughly
for 10 sec. using the vortex mixture.
(6) Place each tube on a magnetic stand, and collect the beads with the magnet.
(7) After magnetic capture, carefully remove the supernatant.
(8) [Washing] Repeat (5) - (7)(9) [Washing] Add 900 μL Washing Solution II, and mix thoroughly for 10 sec.
using the vortex mixture.
(10) Place each tube on a magnetic stand, and collect the beads with the magnet.
(11) After magnetic capture, remove the supernatant carefully.
(12) [Washing] Repeat (9) - (11)(13) [Elution] Add 55 μL Elution Solution, and mix thoroughly for 5 sec.
(14) Heat at 65 °C for 2 min, and mix thoroughly for 5 sec.
(15) Place the tube on a magnetic stand.
(16) Collect the supernatant into a fresh tube.

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.

2NOT FOR HUMAN OR DIAGNOSTIC USE.

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

CauseSolution

Carry over of the magnetic beads inthe Elution solution.
Poor amplification at
RT-PCR
Degradation of RNA[ 6 ] References

Remove the magnetic beads from the Elution
solution. Residual magnetic beads may inhibit
PCR reactions.
Reduce the serum or plasma sample volume forextraction. RNA extracted from excessive
amounts of specimens may contain residual
ribonuclease activity.

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 standa

Code No.
MGS-101

Magical Trapper

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

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

FOR RESEARCH USE ONLY.

3
NOT FOR HUMAN OR DIAGNOSTIC USE.