HPVNALV_Flyer_05171822001:FNR618_LC480_1AK.qxd
12/19/2007
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High Pure Viral Nucleic Acid Large Volume Kit
Expand your viral nucleic acid isolation capabilities
Efficiently isolate total viral nucleic acids from
up to 2.5 ml of mammalian whole blood, serum,
and plasma with the new High Pure Viral Nucleic
Acid Large Volume Kit. Obtain highly purified
viral RNA or DNA using the innovative
combination of a novel extender and proven
High Pure technology (Figure 1).
Use a simple, rapid (less than 25 minutes) protocol
and optimized reagents to generate concentrated,
purified nucleic acids suitable for direct use in
qPCR, qRT-PCR, sequencing, cloning, and other
applications.
쮿
Improve sensitivity.
Use sample volumes up to 2.5 ml to obtain high
yields of purified nucleic acids in a concentrated
50 µl eluate.
쮿
Obtain high-purity nucleic acids.
Reduce carryover risk by using high centrifugal
forces in all wash steps.
쮿
Increase convenience and improve
time to result.
Eliminate complicated sample pre-processing
and rapidly recover purified nucleic acids using
high-speed centrifugation.
Figure 1: High Pure Extender Assembly. The ready-to-use device,
preassembled in a 50 ml polypropylene tube, features an extender that
accommodates large sample volumes. The extender is directly connected
to a High Pure Mini Filter Tube and secured by a locking clip.
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HPVNALV_Flyer_05171822001:FNR618_LC480_1AK.qxd
12/19/2007
9:49 AM
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Fast, efficient viral nucleic acid purification from large sample volumes
Obtain high specificity, outstanding sensitivity, and
excellent linearity in quantitative RT-PCR and PCR
(Figure 2).
쮿
Consistently recover highly pure, concentrated viral
nucleic acids from a broad range of sample sizes,
including volumes as large as 5 ml (Figure 3).
쮿
Produce higher-quality templates compared to other
purification methods (Figure 4).
High Sensitivity
40
30
25
28
26
200 µl
1 ml
2.5 ml
5 ml
24
22
10 6
10 5
10 4
Viral Load (viral equivalents/ml)
Figure 3: Purify nucleic acids from a broad range of sample sizes.
Various amounts (200 µl, 1 ml, 2.5 ml, and 5 ml) of serum research
samples were spiked with a dilution series (1 x 106 to 1 x 104 copies/ml)
of hepatitis A virus (HAV) particles. Nucleic acids were isolated from
each sample using the High Pure Viral Nucleic Acid Large Volume Kit
according to the kit protocol. Five microliters of each sample eluate was
analyzed by LightCycler® qRT-PCR.
Result: The data shows the sensitivity and linearity of the purified
nucleic acids in qRT-PCR analysis, and consistent performance
independent of sample volume. The experiment also demonstrates that
the High Pure Viral Nucleic Acid Large Volume Kit can accommodate
serum sample volumes as large as 5 ml, yielding highly pure,
concentrated nucleic acids.
20
15
10
Performance Comparison
40
5
0
10 5
10 4
10 3
10 2
Viral Load (viral equivalents/ml)
Figure 2: Generate high-quality template with excellent linearity
in qRT-PCR. Citrated plasma research samples were spiked with a
dilution series of hepatitis A virus (HAV) in the range of 102 to 105
copies per ml of plasma. Nucleic acids were isolated from 1 ml of spiked
plasma using the High Pure Viral Nucleic Acid Isolation Large Volume
Kit as described in the kit protocol. Five microliters of eluate from each
isolation was used as template in qRT-PCR analysis with the LightCycler®
Carousel-Based System.
Result: The template purified using the High Pure Viral Nucleic Acid
Isolation Large Volume Kit generated a highly specific PCR product that
demonstrates excellent performance and linearity in qRT-PCR.
Ordering Information
Product.
Product
Pack
Cat. Size
No.
Pack Size
High Pure Viral Nucleic
Acid Large Volume Kit
05 114 403 001
Up to 40 reactions
35
Supplier 1
High Pure Viral Nucleic Acid
Large Volume Kit
30
25
20
10 5
10 6
10 4
HAV Viral Load (viral equivalents/ml)
Performance Comparison
40
Crossing Point Value
Crossing Point Value
35
30
20
Crossing Point Value
쮿
Influence of Sample Size on RT-PCR Performance
32
Crossing Point Value
Overcome the limitations of other viral nucleic acid
isolation techniques. Instead of spending hours to
obtain nucleic acids with less-than-optimal purity and
performance, choose the High Pure Viral Nucleic Acid
Large Volume Kit to generate high-quality, PCR-ready
nucleic acids in less than 25 minutes.
35
Supplier 1
High Pure Viral Nucleic Acid
Large Volume Kit
30
25
20
10 5
10 4
10 6
EBV Viral Load (viral equivalents/ml)
Figure 4: Achieve excellent performance in quantitative
amplification. Serum research samples (1 ml each) were spiked with
different concentrations (106, 105, or 104) of either Epstein-Barr Virus
(EBV) or hepatitis A virus (HAV). Each sample was processed using the
High Pure Viral Nucleic Acid Isolation Large Volume Kit or a kit from
another supplier according to the manufacturers’ protocol. The purified
nucleic acids were then used as templates for qPCR (EBV) or qRT-PCR
(HAV) analysis with the LightCycler® System.
Result: As indicated by the crossing point values, the High Pure Viral
Nucleic Acid Isolation Large Volume Kit provides higher template quality
and sensitivity compared to the kit from another supplier.
Published by
For more information about this kit and other
products for nucleic acid purification, visit
www.roche-applied-science.com/napure
HIGH PURE and LIGHTCYCLER are trademarks of Roche.
Other brands or product names are trademarks of their respective holders.
Roche Diagnostics GmbH
Roche Applied Science
68298 Mannheim
Germany
© 2007 Roche Diagnostics
05171822001햲1207