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BR-8134A
P/ACETM DNA SYSTEM Flexible, Automated DNA Analysis
BECKMAN
•
A CLEAR ADVANTAGE
FOR MUTATION ANALYSIS,
POLYMORPHISMS,
SYNTHETIC OLIGONUCLEOTIDES,
DNA QUANTITATION,
AND PROTEIN/DNA
INTERACTION
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I HE P/ACE DNA SYSTEM COMBINES TO 61.
THE VISIBLE BENEFITS OF CE FOR DNA ANALYSIS
Just as DNA is not complete without all
four nucleotides, a total capillary electro-phoresis (CE) solution for your application
requires the right mix of hardware, soft-
ware, chemistry and support. The P/ACE
DNA System combines the power of the
P/ACE 5000 capillary electrophoresis
system, LIF (laser-induced fluorescence) detection, P/ACE software, and a broad
range of eCAP'" chemistries with the most
experienced CE support team in the world. The result: automated DNA analysis with
a clarity that is unmatched by traditional
gel electrophoresis methods.
AUTOMATION With the need to do more with less, the P/ACE DNA System
easily automates the DNA analysis
process from gel preparation to data han-
dling. You will save time and money while obtaining better reproducibility and more
information than with slab gels. The
P/ACE 5000's automated sample handling
capabilities eliminate the need for manual
sample loading and gel preparation.
And with on-capillary detection,
quantitation is automatic. There is no time
wasted staining, reading gels with a scanner
or disposing of hazardous materials.
With Beckman's seven years of CE DNA separation experience you spend less
time working out instrument and method
conditions and more time understanding your DNA.
EPRESENTATIVE APPLICATION AREAS OF THE P/ACE DNA SYSTEM
CATEGORY APPLICATION REFERENCE
SIZE POLYMORPHISMS VNTR
STR (microsatellites)
RFLP
RAPD
Beckman Primer 607397
Ulfelder, Kathi and Christian Oste, Sixth Annual Frederick Conference on Capillary Electrophoresis, Session 8 (1995)
'Ulfelder, Kathi et al., Analytical Biochemistry 200 (1992): 260-267
*Tattersall, David et al., Australian Cooperative Research Centre for Viticulture, Program 1, Subprogram 1.1
DNA QUANTITATION Viral Load Determination
Competitive RT-PCR' to measure gene expression
Plasmid Analysis
Schartz, Herbert et al., Journal of Capillary Electrophoresis 1 (1994): 36-54
Beckman Application Information Bulletin A-1788A
Courtney, Bernard et al., Analytical Biochemistry 228 (1995): 281-286
MUTATION ANALYSIS SSCP 'Landers, James et al., Seventh International Symposium on High Performance Capillary Electrophoresis. Poster 313 (1995)
SYNTHETIC OLIGONUCLEOTIDES Purity Analysis 'Beckman Bulletin 7912 Beckman Technical Information Bulletin T-1794A
PROTEIN/DNA INTERACTION Mobility Shift Assay Capillary Electrophoresis. Poster TI21 (1993)
Waschke, Hans et al., Fifth International Symposium on High Performance
DNA quantitation, mutation detection, size polymorphism analysis and
separation of synthetic oligonucleotides are quickly performed using the
automated P/ACE DNA System.
' UV detection was used in the listed reference
' The Polymerase Chain Reaction (PCR) is covered by U.S. Patents owned by Hoffman-LaRoche Inc. All trademarks are the property of their respective owners.
The P/ACE DNA System automates
the separation, quantitation, analysis
and documentation of oligonucleotides,
Polymerase Chain Reaction (KR)
products, and restriction fragments.
Beckman's broad range of eCAP
chemistries, combined with the powerful
P/ACE DNA system, automatically
speeds every aspect of DNA analysis.
CAP NA 110-N Kit
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YOU A CLEAR ADVANTAGE IN DNA SEPARATIONS Iso .0 n 01.V. 444 * 000000000 b 440 4 0 * 0 404 440*•44i4 3 44 044. 4". 4g4.3.
BECKMAN'S BROAD RANGE OF eCAP CHEMISTRIES, COMBINED WITH THE POWERFUL P/ACE DNA SYSTEM, AUTOMATICALLY SPEEDS EVERY ASPECT OF DNA ANALYSIS
CLARITY Seeing the "big" picture is
one of the main benefits of the P/ACE
DNA System. From fragment analysis to
DNA-Protein Interaction, you will be
able to separate and quantify with con-
tidence. High resolution separations are
possible with the eCAP DNA capillary
and its replaceable, polyacrylamide gel.
Automated replacement minimizes contam-ination and results in excellent run-to-run
reproducibilities. Combining the selectivity
and sensitivity of the LIF (laser-induced
fluorescence) detector with the data
collection and integration capabilities of
the P/ACE software you can be assured
of seeing minor variances, while using
only a few nanoliters of sample.
As shown in the table (below
left) and on the following pages, the wide
variety of separations possible with the
P/ACE DNA System proves that it is the right combination of CE hardware, soft-
ware, chemistry and support for your
DNA application.
MMEDIATE RESULTS IN A VARIETY OF EV EE
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T24 1000 by
200 by T18 T24
T14 T311000 by
200 by
D1580 PCR products
26 28 30 32 34 36 38 40 42 44 46 TIME (minutes)
I10 RFU
D1S80 Allelic ladder
eCAP dsDNA 1000 Kit
Calibration curve for molecular size
determination. Curve was generated
using dsDNA fragments from a
oX 174 DNA/Fir/ell! digest separated
with the eCAP dsDNA 1000 Kit.
2.986Correlalion Coefficient 0991275
Linear
B
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VNTR analysis of the PCR-amplified
D1580 locus from a heterozygous
individual, compared to the allelic
standard using the eCAP dsDNA 1000
Kit. A 32 by difference between allelic
fragments is demonstrated in the
sample. Standards of 200 and
1000 by are added for sizing the
DNA fragments.
STR analysis of PCR-amplified genetic
loci, THOI, TPDX, and FES/FPS from
three individuals in one family, demon-
strating paternity. Separations using
the eCAP ssDNA 100-R Kit with
EnhanCE interculator and LIF detection
detect differences of 4 by in size.
TH01 TPDXFES/FPS
Mother
Child
Father
J 22 26 30
34
38 TIME (minutes)
IZE POLYMORPHISMS
Size polymorphism analysis is used to
correlate polymorphic sequences of DNA
with a gene of interest. The P/ACE DNA System with the eCAP T" dsDNA 1000
Kit and EnhanCE intercalating dye pro-vides fast runs with high resolution for PCR-amplified restriction fragment length
polymorphism (PCR-RFLP) or random amplified polymorphic DNA PCR (RAPD-PCR) analyses. LIF detection
provides the high sensitivity required for low sample levels.
DNA profiling or typing for paternity testing and forensic applications
can be done with short tandem repeat (STR) or variable number tandem repeat. (VNTR) analyses. Good reproducibility
and software for molecular sizing facilitates
pattern recognition.
14 15 16
18 TIME (minutes)
Sabin 3
primers and primerAimer
w 7 '2)
Ew 5-
ST
C
3
100CF
80TARGET
0 COMPETITOR 60
U N3 cci
LL20 Is
20
30 TIME (minutes)
Intact plasmid
110 RFU
25 21 13 17 TIME (minutes)
EcoRl full digest
3 Kbp inert —
12 Kbp
9 Kbp vector
CE-LIF analysis of DNA products
generated by competitive RNA-PCR
of cytochrome P450-1A1 mRNA in
dioxin-treated HepG2 cells. Target
sequence is 367 bp; competitor is
228 bp; CR = 5-carboxyfluorescein
marker; IS = 328 by internal standard.
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L.ATIONS 911404 OOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOO 0000.011011.03.i44G00074.401.89.04.11.4640071.004.4408 66666666666666666666666666666666666666666666 666666666666666 leo
"Before and after" separation of
DNA fragments derived from the
complete EcoRl digestion of a 12 Kb
plasmid vs. the undigested plasmid
itself using LIF detection.
LIF (laser-induced fluorescence)
detection of the separation of a
53 by RT-PCR product from the RNA
of the Sabin-3 strain of the polio virus
vaccine. A Haelll-digested oX 174
DNA marker, at a concentration of
10 pg/mL, was co-injected with the
PCR product for size verification.
NA QUANTITATION
Polymerase Chain Reaction (PCR) with
the electrophoretic separation of the amplified products provides the ability to detect and quantitate small amounts
of target DNA. The P/ACE DNA System automates the analysis of the PCR prod-
ucts. With the unique co-injection feature, size and/or quantitation standards are
loaded automatically to eliminate the need for pre-mixing with each sample.
Accurate quantitation at low
levels can often be best achieved through competitive PCR techniques such as
competitive RNA-PCR. The P/ACE DNA System provides superior precision
and eliminates the need for off-line
detection and scanning. Applications such as monitoring
plasmid copy number in cell cultures
require quantitation of large DNA frag-
ments up to 20,000 base pairs. The eCAP'" dsDNA 20,000 Kit includes the capillary, gel, and size standards for these applications.
The analysis of large DNA fragments
can also be used for monitoring plasmid
ligation efficiency where the efficiency is maximized by the ultra-low
sample consumption of the P/ACE DNA System.
eCAP dsDNA 20,000 Kit
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FLEXIBILITY TO ADAPT TO YOUR CHANGING
23 25
Trityl-off
17 19 21 TIME (minutes)
Tvityl-on
0.036
0.034
0.030
0.026
0.022
48 0.018
Zi 0.014 0
aim
0.006-
0.002-
-0.002 15
Packaging - P _
Purity analysis of a crude synthetic
100-mer using eCAP ssDNA 100-R Kit
and UV Detector Option.
Separation of a synthetic 17-mer
oligonucleotide with the trityl group
attached to the 5' end, performed
using the eCAP ssDNA 100-R Kit and
the UV Detector Option. Inset shows
high-speed separation of the same
oligonucleotide accomplished using the
7cm portion of the capillary.
Synthetic 21-mer that is fluorescein-
labeled and purified following synthesis
using the eCAP ssDNA 100-R Kit and
the LIF (laser-induced fluorescence)
detector with the 488 Laser Module.
Flexibility means having a system that can
adapt to meet your varied needs in DNA analysis.
UV detection is available for applications such
as quality checking of synthetic oligonucleotides
while additional laser sources such as the 635 Laser Module allow you to choose the most
convenient or best performing fluorescent dye
for your application. To allow the analysis of
small oligonucleotides and nucleic acids, the
P/ACE DNA System is ideal for non-gel
methods such as capillary zone electrophoresis
(CZE) or micellar electrokinetic capillary
chromatography (MECC).
YNTHETIC OLIGONUCLEOTIDES
0 For applications that are tedious or involve
precious, expensive samples, the quality of (—
oligonucleotide primers and probes is critical. The P/ACE DNA system with the eCAP
ssDNA 100-R Kit provides a fully automated,
rapid, and high-resolution system for checking
these oligonucleotides. Labeled oligonucleotides
may be analyzed using LIF detection. UV
detection is available for the analysis of unlabeled
oligonucleotides including antisense phospho-
rothioate oligonucleotides.
eCAP ssDNA 100-R Kit
OLIO co es oin
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1000" ...... 00000000%0 OOOOOOOOOOOOO 0 OOOOOOOOO 0000 OOOOO 00000000 OOOOOOOOOO 0000000000000 OOOOOOOOOOOOOOOOOOOOOOOOOOO 00000000000000000000000000 OOOOOOOOOO 0000
SSCP analysis of PCR-amplified rpoB
gene of M. Tuberculosis indicative of
rifampicin susceptibility (UV detection).
Separation of single-base mutant
strands is shown. Data provided
courtesy of Dr. James Landers,
Director of the Clinical CE facility
at the Mayo Clinic.
Binding of EcoRl enzyme to a JOE-
labeled oligonucleotide. Unbound DNA
versus EcoRl complexed with DNA
are compared. Data provided courtesy
of Dr. P. Mickey Williams, Genentech.
LIF detection with 488 laser and
540 emission.
P/ACE DNA System with
optional 635 Laser Module
Capillary electrophoresis is an excellent
tool for DNA analysis, and the application
of this technology continues to grow
rapidly. With Beckman, the world leader
in capillary electrophoresis, you will keep
pace with these continuing developments.
OUTATION ANALYSIS
Methods for detecting mutations in DNA often require unique electrophoretic con-
ditions. One technique, single-strand
conformation polymorphism (SSCP)
analysis, is based on mobility differences
of ssDNA fragments in non-denaturing gels caused by conformational differences due to point mutations.
ROTEIN/DNA INTERACTION
Proteins and their molecular interactions with DNA are important areas of study in
molecular biology. Protein binding is easily monitored through mobility shift assays.
DNA APPLICATION BULLETINS
APPLICATION BULLETINS
A-1748A
Capillary Electrophoresis of dsDNA Fragments with UV and Laser-Induced Fluorescence Detection
A-1774A
Quantitative Capillary Electrophoretic Analysis of PCR Products Using Laser-Induced Fluorescence Detection
A-1788A
Competitive RNA-PCR by Capillary Electrophoresis and Laser-Induced Fluorescence (LIF) Detection for Quantitation of Cellular mRNA
DS-738A
Analysis of Nucleic Acid Derivatives and Analogs by P/ACE Using Capillary Electrophoresis
SR-183
High Sensitivity Analysis of Nucleotides Using Electrokinetic Injection and Sample Stacking with Micellar Electrokinetic Capillary Chromatography (MECC)
T- 1779A Purity Assessment of Two Synthetic Oligonucleotide Preparations by Capillary Gel Electrophoresis with the P/ACE System
OoomegOAINNIMMIMI
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SUPPORT
SOLUTIONS BECKMAN I.S. EN ISO 9001
Ordering Information
P/ACE DNA SYSTEMS
267817 P/ACE DNA System, LIF detection only (120V). Includes P/ACE DNA System with LIF detector, 488 Laser Module, Workstation Software, LIF Calibration/Test Standard, eCAP dsDNA 1000 and 20,000 Kits, EnhanCE Intercalator, and Literature Package.
261844 P/ACE DNA System, LIF detection only (240V)
267819 P/ACE DNA System, LIF detection only (220V)
267820 P/ACE DNA System, LIF detection only (100V)
267818 P/ACE DNA System, LIF and UV detection (120V). Includes P/ACE DNA System with LIF and UV detectors, 488 Laser Module, Workstation Software, LIF Calibration/Test Standard, eCAP dsDNA 1000 and 20,000 Kits, EnhanCE Intercalator, and Literature Package.
267847 P/ACE DNA System, LW and UV detection (240V)
267845 P/ACE DNA System, LIF and UV detection (220V)
261846 P/ACE DNA System, LIF and UV detection (100V)
ACCESSORIES
267848 UV Detector Upgrade for LIF-only Systems. Includes the UV Detector Options, UV Detector Head and Filters. Field service installation is also included.
267839 635 Laser Module. Includes Semiconductor 635 nm Laser, Calibration/Test Standard, 675 nm Emission Filter, Extra Filter Holder.
DNA CHEMISTRY KITS
477410 eCAP dsDNA 1000 Kit. For analysis of double-stranded DNA with linearity from 100 to 1,000 base pairs. Includes coated capillaries (2), replaceable gel, and standards to perform at least 100 runs.
471486 eCAP dsDNA 20,000 Kit. For analysis of double-stranded DNA with linearity from 1,000 to 20,000 base pairs. Includes coated capillaries (2), replaceable gel, and standards to perform at least 100 runs.
471480 eCAP ssDNA 100-R Kit. For analysis of single-stranded DNA with linearity from 10 to 100 bases. Includes coated capillaries (2), replaceable gel, and standards to perform at least 100 runs. UV detection is required to analyze unlabeled and standard oligonucleotides.
477409 EnhanCE Intercalator, 0.5 ml. For use with either eCAP dsDNA 1000 or eCAP dsDNA 20,000 Kits.
T 11 • E
BECKMAN
Beckman Instruments, Inc. • 2500 Harbor Boulevard, Box 3100 • Fullerton, California 92634-3100 Sales: 1-800-742-2345 • Service: 1-800-551-1150 • Internet: http://www.beckman.com • Telex: 678413 • Fax: 1-800-643-4366
Worldwide Offices: Africa, Middle East, Eastern Europe (Switzerland) (41) 22 994 07 07. Australia (61) 2 844 6000. Austria (43) 222 7292164. Canada (800) 387-6799. China (861) 5051241-2. France (33) 1 43 01 70 00. Germany (49) 89-358700. Hong Kong, PRC (852) 814 7431. Italy (39) 2-953921. Japan 3-5352-2825. Mexico (525) 559-1635. Netherlands 0297-230630. Poland (48) 22 408822, 408833. Singapore (65) 339 3633. South Africa (27) 11-805-2014/5. Spain (1) 358-0051. Sweden (8) 445 11 20. Switzerland (41) 22 994 07 07. Taiwan (886) 02 378-3456. U.K. (01494) 441181. U.S.A. 1-800-742-2345.
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©1996 Beckman Instruments, Inc. 0 Printed in U.S.A. on recycled paper.