August 2014
SpeedXtractTM Nucleic Acid Kit
Handbook
For the rapid manual extraction of nucleic
acids from a wide range of biological
samples
Sample & Assay Technologies
QIAGEN Sample and Assay Technologies
QIAGEN is the leading provider of innovative sample and assay technologies,
enabling the isolation and detection of contents of any biological sample. Our
advanced, high-quality products and services ensure success from sample to
result.
QIAGEN sets standards in:

Purification of DNA, RNA, and proteins

Nucleic acid and protein assays

microRNA research and RNAi

Automation of sample and assay technologies
Our mission is to enable you to achieve outstanding success and breakthroughs.
For more information, visit www.qiagen.com.
Contents
Kit Contents
4
Storage
4
Product Use
4
Safety Information
4
Quality Control
4
Introduction
5
Principle and procedure
5
Description of protocols
7
Equipment and Reagents to Be Supplied by User
Important Notes
9
10
Starting material
10
Lysis efficiency
10
Nucleic acid yields
10
Nucleic acid purity
10
Protocols:
 SpeedXtract Nucleic Acid Extraction from Swab Samples
12
 SpeedXtract Nucleic Acid Extraction from Liquid Samples
13
 SpeedXtract Nucleic Acid Extraction from Complex Liquid Samples
15
Troubleshooting Guide
17
Ordering Information
19
SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
3
Kit Contents
SpeedXtract Nucleic Acid Kit
(200)
Catalog no.
703060
Number of preparations
200
Buffer EN
2 x 150 ml
Buffer SL
2 x 100 ml
SpeedXtract Suspension A
4 x 1.5 ml
QuickStart Protocol
1
Storage
The SpeedXtract Kit (cat. no. 703060) can be stored at room temperature (15–
25°C) for up to 12 months.
Product Use
For Research Use Only. Not for use in diagnostic procedures.
No claim or representation is intended to provide information for the diagnosis,
prevention, or treatment of a disease. The performance characteristics of this
product have not been fully established.
All due care and attention should be exercised in the handling of all products.
The SpeedXtract Nucleic Acid Kit is intended for rapid manual extraction of
crude nucleic acids from a wide range of biological samples.
Safety Information
When working with chemicals, always wear a suitable lab coat, disposable
gloves, and protective goggles. For more information, please consult the
appropriate safety data sheets (SDSs). These are available online in convenient
and compact PDF format at www.qiagen.com/safety where you can find, view,
and print the SDS for each QIAGEN kit and kit component.
Quality Control
In accordance with QIAGEN’s ISO-certified Quality Management System, each
lot of SpeedXtract Nucleic Acid Kit is tested against predetermined
specifications to ensure consistent product quality.
4
SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
Introduction
The SpeedXtract Nucleic Acid Kit provides a fast and easy way of extracting
crude nucleic acids from biological samples without the need for a sophisticated
laboratory environment. The kit can be used with a variety of sample types,
including swabs, pre-concentrated cells, and various liquid samples, such as
urine or blood.
The nucleic acid extracts are ready for use in downstream applications such as
amplification of viral or bacterial nucleic acids using PCR or isothermal
amplification methods.
Principle and procedure
The SpeedXtract Nucleic Acid Kit is designed for rapid manual extraction of
nucleic acids using efficient heat lysis and convenient separation using
suspended magnetic particles (see flowchart for swab and liquid samples, page
6).
Swab samples are eluted in the SpeedXtract extraction buffer (Buffer SL) and a
suspension of magnetic particles (SpeedXtract Suspension A) is added. Heat
lysis at 95°C efficiently lyses multiple cell types, including eukaryotic cells,
viruses, and bacteria. Nucleic acids are released into Buffer SL, and cell debris,
particulate matter, and contaminants are bound to the suspended magnetic
particles. The magnetic particles are then easily separated using a magnetic
stand, and the supernatant containing the nucleic acids can be used directly in
downstream applications.
Liquid samples require analyte enrichment prior to nucleic acid extraction. The
samples are mixed with the SpeedXtract enrichment buffer (Buffer EN) and
SpeedXtract Suspension A. The magnetic particles in SpeedXtract Suspension A
bind to biological targets, including eukaryotic cells, viruses, and bacteria. After
the magnetic particles are separated and the supernatant is removed, the
magnetic particles and attached cells can be resuspended directly in Buffer SL.
Nucleic acid extraction then proceeds as described above.
SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
5
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SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
Description of protocols
Swab samples can undergo cell lysis and rapid nucleic acid extraction
immediately whereas liquid samples, such as urine, saliva, serum, and plasma,
require analyte enrichment prior to nucleic acid extraction. For complex liquid
matrices, such as whole blood, a wash step following analyte enrichment and
before nucleic acid extraction, is recommended. Performance is not guaranteed
for every combination of starting material and cell type and must be validated
by the user after consideration of the “Important Notes”, page 10.
SpeedXtract nucleic acid extraction from swab samples (page 12)
Swab samples are eluted by swirling in 200 µl – 1 ml of the SpeedXtract
extraction buffer (Buffer SL). For more information on choosing an elution
volume, see “Starting material”, page 10. A suspension of magnetic particles
(SpeedXtract Suspension A) is then added. Heat lysis releases nucleic acids into
Buffer SL, whereas cell debris, particulate matter, and contaminants are bound
to the suspended magnetic particles. The magnetic particles are then separated
using a magnetic stand, and the supernatant containing the nucleic acid extract
can be used directly in downstream applications.
SpeedXtract nucleic acid extraction from liquid samples (page 13)
For liquid samples, such as urine and saliva, 200 µl – 1 ml of sample is mixed
with the same volume of the SpeedXtract enrichment buffer (Buffer EN). A
suspension of magnetic particles (SpeedXtract Suspension A) is then added. The
magnetic particles in SpeedXtract Suspension A bind to biological target
material, including eukaryotic cells, viruses, and bacteria. The magnetic
particles are then separated using a magnetic stand, and the supernatant is
removed. The magnetic particles and attached cells are resuspended in 100 –
200 µl of Buffer SL, depending on the amount of sample material. Nucleic acid
extraction then proceeds as described for swab samples.
SpeedXtract nucleic acid extraction from complex liquid samples (page 15)
For complex liquid matrices, such as whole blood, up to 200 µl of sample is
mixed with 2 volumes of the SpeedXtract enrichment buffer (Buffer EN). A
suspension of magnetic particles (SpeedXtract Suspension A) is then added. The
magnetic particles in SpeedXtract Suspension A bind to biological target
material, including eukaryotic cells, viruses, and bacteria. The magnetic
particles are then separated using a magnetic stand, and the supernatant is
removed. The magnetic particles and attached cells are washed by
resuspending in Buffer EN, with magnetic particle separation and removal of the
SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
7
supernatant as described above. The magnetic particles and attached cells are
then resuspended in 100 – 200 µl of Buffer SL, depending on the amount of
sample material. Nucleic acid extraction then proceeds as described for swab
samples.
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SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
Equipment and Reagents to Be Supplied by User
When working with chemicals, always wear a suitable lab coat, disposable
gloves, and protective goggles. For more information, consult the appropriate
safety data sheets (SDSs), available from the product supplier.

Pipets and disposable pipet tips with aerosol barriers

2 ml microcentrifuge tubes with screw caps or lock caps

1.5 ml microcentrifuge tubes

Magnetic stand, e.g. QIAGEN MagAttract® Magnetic Rack (cat. no.
19606)

Vortexer

Water bath, heating block, or thermal shaker (e.g. Eppendorf®
Thermomixer) for 2 ml microcentrifuge tubes capable of holding
temperatures up to 95°C
SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
9
Important Notes
Starting material
Swab samples are directly eluted in 200 µl – 1 ml Buffer SL before proceeding
with the SpeedXtract nucleic acid extraction procedure. Elution of swabs in low
volumes generally increases the concentration of target material but also
increases the concentration of potential inhibitors. This can lead to impaired
extraction of nucleic acids and/or performance in downstream assays. Use
appropriate controls (e.g. an internal control) to verify successful amplification.
When choosing an elution volume, consider that swabs retain different amounts
of liquid depending on their composition; for example, woven swabs retain
more liquid than flocked swabs.
For liquid samples, enrichment of the target analyte is recommended prior to
proceeding with the SpeedXtract nucleic acid extraction procedure. Liquid
samples are mixed with the SpeedXtract enrichment buffer (Buffer EN). For most
liquid matrices, including urine and saliva, equal volumes of Buffer EN and
sample are recommended. For complex liquid matrices, such as whole blood,
2 volumes of Buffer EN are mixed with one volume of sample. A further wash
step with Buffer EN is also recommended for complex liquid matrices.
Lysis efficiency
For enhanced lysis efficiency, especially for viruses and eukaryotic cells, heat
incubation can be extended to up to 15 minutes. Additionally, shaking during
heat incubation can be used to improve heat transfer into the lysis solution.
Nucleic acid yields
The SpeedXtract Nucleic Acid Kit recovers total nucleic acids, including both
cellular and viral DNA and RNA (although extraction efficiency for RNA is low).
The DNA derived from these different sources cannot be distinguished by
spectrophotometric analysis. In addition, in samples with low numbers of cells,
such as urine, nucleic acid yields may be less than 1 µg. Therefore, it is
recommended to use quantitative real-time PCR for determining yield of the
primary nucleic acid target.
Nucleic acid purity
Extracted nucleic acids may contain small amounts of contaminants, such as
urea, heme, or proteins, depending on the sample type. Therefore, the crude
nucleic acid extracts that have been obtained with the SpeedXtract Nucleic Acid
10
SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
Kit are suitable only for downstream applications that do not require highly pure
nucleic acids, such as PCR or isothermal amplification. Nucleic acid extracts
that have been obtained with the SpeedXtract Nucleic Acid Kit are not
recommended for use in downstream applications that require highly pure
nucleic acids, such as DNA sequencing.
SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
11
Protocol: SpeedXtract Nucleic Acid Extraction from Swab
Samples
This protocol is for the rapid manual extraction of crude nucleic acids from swab
samples (e.g. nasal, throat or buccal swabs).
Important points before starting

Use 2 ml microcentrifuge tubes with screw caps or lock caps to avoid
accidental opening during heat lysis
Things to do before starting

Heat a water bath, heating block, or thermal shaker for 2 ml
microcentrifuge tubes to 95°C
Procedure
1. Elute swabbed material by swirling in 200 µl – 1 ml of Buffer SL in a 2 ml
microcentrifuge tube (not provided).
The elution volume can be altered depending on the amount of sample
material. Elution of swabs in low volumes generally increases the
concentration of target material but also increases the concentration of
potential inhibitors.
2. Vortex SpeedXtract Suspension A for 30 seconds to resuspend magnetic
particles. Add 30 µl of resuspended SpeedXtract Suspension A to the sample
tube. Mix well for 10 seconds by vortexing or inverting.
3. Incubate sample tubes at 95°C for 5 minutes.
If a thermal shaker is used, shake at maximum speed.
Note: For enhanced lysis efficiency, especially for viruses and eukaryotic
cells, incubate samples at 95°C in a thermal shaker at maximum speed for
up to 15 minutes.
4. Remove sample tubes from heat source and either shake down or tap on a
bench to remove condensate from the lid.
5. Transfer sample tubes into a magnetic stand and incubate at room
temperature for 1 minute.
6. Carefully open sample tubes and transfer supernatant (i.e. nucleic acid
extract) to a new tube (not provided) for storage or use an aliquot directly in
applications (e.g. PCR or isothermal amplification).
Nucleic acid extracts can be stored at −20oC.
12
SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
Protocol: SpeedXtract Nucleic Acid Extraction from Liquid
Samples
This protocol is for the rapid manual extraction of crude nucleic acids from
liquid samples (e.g. urine, saliva, serum, or plasma) and includes an analyte
enrichment step prior to nucleic acid extraction.
Important points before starting

Use 2 ml microcentrifuge tubes with screw caps or lock caps to avoid
accidental opening during heat lysis
Things to do before starting

Heat a water bath, heating block, or thermal shaker for 2 ml
microcentrifuge tubes to 95°C
Procedure
1. Add 200 µl – 1 ml of liquid sample to a 2 ml microcentrifuge tube.
2. Add the same volume of Buffer EN as the volume of sample used in step 1.
3. Vortex SpeedXtract Suspension A for 30 seconds to resuspend magnetic
particles. Add 30 µl of resuspended SpeedXtract Suspension A to the sample
tube. Mix well for 10 seconds by vortexing or inverting. Shake down or tap
on a bench to remove any liquid from the lid.
4. Incubate sample tubes at room temperature for 3 minutes.
5. Transfer sample tubes into a magnetic stand and incubate at room
temperature for 1 minute.
6. Carefully open sample tubes and remove the supernatant without disturbing
the magnetic particle pellet. Discard the supernatant.
7. Add 100 – 200 µl of Buffer SL to sample tubes and resuspend the magnetic
particles by vortexing or pipetting.
The elution volume can be altered depending on the amount of sample
material. A lower volume of Buffer SL may increase the concentration of
nucleic acids in the extract.
8. Incubate sample tubes at 95°C for 5 minutes.
If a thermal shaker is used, shake at maximum speed.
Note: For enhanced lysis efficiency, especially for viruses and eukaryotic
cells, incubate samples at 95°C in a thermal shaker at maximum speed for
up to 15 minutes.
SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
13
9. Remove sample tubes from heat source and either shake down or tap on a
bench to remove condensate from the lid.
10. Transfer sample tubes into a magnetic stand and incubate at room
temperature for 1 minute.
11. Carefully open sample tubes and transfer supernatant (i.e. nucleic acid
extract) to a new tube (not provided) for storage or use an aliquot directly in
applications (e.g. PCR or isothermal amplification).
Nucleic acid extracts can be stored at −20oC.
14
SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
Protocol: SpeedXtract Nucleic Acid Extraction from
Complex Liquid Samples
This protocol is for extraction of crude nucleic acids from whole blood and other
complex liquid matrices, and includes an analyte enrichment step and a
subsequent wash step prior to nucleic acid extraction.
Important points before starting

Use 2 ml microcentrifuge tubes with screw caps or lock caps to avoid
accidental opening during heat lysis
Things to do before starting

Heat a water bath, heating block, or thermal shaker for 2 ml
microcentrifuge tubes to 95°C
Procedure
1. Add up to 200 µl of liquid sample to a 2 ml microcentrifuge tube.
2. Add 2 volumes of Buffer EN to 1 volume of the liquid sample used in step 1.
3. Vortex SpeedXtract Suspension A for 30 seconds to resuspend magnetic
particles. Add 30 µl of resuspended SpeedXtract Suspension A to the sample
tube. Mix well for 10 seconds by vortexing or inverting. Shake down or tap
on a bench to remove any liquid from the lid.
4. Incubate sample tubes at room temperature for 3 minutes.
5. Transfer sample tubes into a magnetic stand and incubate at room
temperature for 1 minute.
6. Carefully open sample tubes and remove the supernatant without disturbing
the magnetic particle pellet. Discard the supernatant.
7. Add 500 µl of Buffer EN and resuspend magnetic particles by vortexing or
pipetting. Transfer sample tubes into a magnetic stand and incubate at room
temperature for 1 minute. Carefully open sample tubes and remove the
supernatant without disturbing the magnetic particle pellet.
8. Add 100 – 200 µl of Buffer SL to the sample tubes and resuspend the
magnetic particles by vortexing or pipetting.
The elution volume can be altered depending on the amount of sample
material. A lower volume of Buffer SL may increase the concentration of
nucleic acids in the extract.
9. Incubate sample tubes at 95°C for 5 minutes.
If a thermal shaker is used, shake at maximum speed.
SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
15
Note: For enhanced lysis efficiency, especially for viruses and eukaryotic
cells, incubate samples at 95°C in a thermal shaker at maximum speed for
up to 15 minutes.
10. Remove sample tubes from heat source and either shake down or tap on a
bench to remove condensate from the lid.
11. Transfer sample tubes into a magnetic stand and incubate at room
temperature for 1 minute.
12. Carefully open sample tubes and transfer supernatant (i.e. nucleic acid
extract) to a new tube (not provided) for storage or use an aliquot directly in
applications (e.g. PCR or isothermal amplification).
Nucleic acid extracts can be stored at −20oC
16
SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
Troubleshooting Guide
This troubleshooting guide may be helpful in solving any problems that may
arise. For more information, see also the Frequently Asked Questions page at
our Technical Support Center: www.qiagen.com/FAQ/FAQList.aspx. The
scientists in QIAGEN Technical Services are always happy to answer any
questions you may have about either the information and protocols in this
handbook or sample and assay technologies (for contact information, see back
cover or visit www.qiagen.com).
Comments and suggestions
Nucleic acids do not perform well in downstream applications
a) Carryover of salts or
inhibitory substances
during extraction
After magnetic particle separation, ensure that
the supernatant is removed carefully without
disturbing the magnetic particle pellet.
If samples were prepared according to a
protocol without an analyte enrichment step and
a wash step before nucleic acid extraction, use
the protocol for complex liquids, which includes
an analyte enrichment step and a wash step.
If the nucleic acid extract still contains inhibitory
substances, dilute the extract before use in
downstream applications.
b) Insufficient nucleic
acids for use in
downstream
application
If samples were prepared according to a
protocol without an analyte enrichment step, use
the protocol for liquids, which includes an
analyte enrichment step.
Ensure SpeedXtract Suspension A is fully
resuspended before use.
If samples were prepared according to a
protocol with an analyte enrichment step,
increase the ratio of Buffer EN added to the
sample, e.g. from 1:1 (i.e. 1 volume of Buffer EN
to 1 volume of sample) to 2:1 or 3:1.
SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
17
Comments and suggestions
c) Insufficient lysis
Extend the heat incubation from 5 minutes
stepwise to 15 minutes.
If a water bath was used, use a thermal shaker at
maximum speed.
If samples were prepared according to a
protocol with an analyte enrichment step, ensure
all supernatant is removed before resuspending
the magnetic particles in Buffer SL.
d) Magnetic particle
carryover
18
Handle sample tubes with care after separation
of magnetic particles taking care to not disturb
the magnetic particle pellet. If carryover does
occur, magnetic particles in extracts will not
affect most downstream applications.
SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
Ordering Information
Product
Contents
Cat. no.
SpeedXtract Nucleic
Acid Kit
For 200 preps: Buffers, and
SpeedXtract Suspension A
703060
Accessories
MagAttract Magnetic
Rack
Magnetic rack for convenient parallel
processing of up to 12 samples
19606
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SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
19
Notes
20
SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
Notes
SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
21
Notes
22
SpeedXtract™ Nucleic Acid Kit Handbook 08/2014
Trademarks: QIAGEN®, MagAttract®, SpeedXtract™ (QIAGEN Group); Eppendorf®, Thermomixer® (Eppendorf AG).
Limited License Agreement for SpeedXtract Nucleic Acid Kit
Use of this product signifies the agreement of any purchaser or user of the product to the following terms:
1.
The product may be used solely in accordance with the protocols provided with the product and this handbook and for use with components
contained in the kit only. QIAGEN grants no license under any of its intellectual property to use or incorporate the enclosed components of this
kit with any components not included within this kit except as described in the protocols provided with the product, this handbook, and
additional protocols available at www.qiagen.com. Some of these additional protocols have been provided by QIAGEN users for QIAGEN
users. These protocols have not been thoroughly tested or optimized by QIAGEN. QIAGEN neither guarantees them nor warrants that they do
not infringe the rights of third-parties.
2.
Other than expressly stated licenses, QIAGEN makes no warranty that this kit and/or its use(s) do not infringe the rights of third-parties.
3.
This kit and its components are licensed for one-time use and may not be reused, refurbished, or resold.
4.
QIAGEN specifically disclaims any other licenses, expressed or implied other than those expressly stated.
5.
The purchaser and user of the kit agree not to take or permit anyone else to take any steps that could lead to or facilitate any acts prohibited
above. QIAGEN may enforce the prohibitions of this Limited License Agreement in any Court, and shall recover all its investigative and Court
costs, including attorney fees, in any action to enforce this Limited License Agreement or any of its intellectual property rights relating to the kit
and/or its components.
For updated license terms, see www.qiagen.com.
© 2014 QIAGEN, all rights reserved.
www.qiagen.com
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1083147
08/2014
Sample & Assay Technologies