Nucleic Acid Amplification
AmpliGrid System
1µL Assays
Nucleic Acid Amplification – Highest sensitivity down to ONE single cell
2
AMPLIGRID SYSTEM – INTRODUCTION
SETTING NEW STANDARDS
The art of downscaling biological reactions
To date, researchers focusing on the molecular biology of single cells rely on microplates or tubes
for their amplification assays. As a result, large dead volumes, template absorption and no optical
quality control are responsible for a considerable uncertainty and low reproducibility. Furthermore,
the average sensitivity of microplate-based systems is usually far below the user’s expectations.
With the innovative AmpliGrid system from Advalytix, sensitivity, efficiency and reproducibility are
reliable constants for both single cell and ultra low volume amplification techniques. And as a result
fast and convenient workflow is assured.
Advalytix is introducing a superior amplification platform based on a chemically structured
microscope slide. The AmpliGrid system is the key to obtaining perfect results especially with
single cells and nearly any other kind of template material e.g., forensic stains or purified nucleic
acids. Due to the two-dimensional layout of the AmpliGrid, single cells can easily be deposited
to the slide surface, e.g. by a fluorescence activated cell sorter (FACS), micromanipulator or
microdissection, rendering a variety of single cell applications possible as well as providing full
access to the “one-cell genome”.
The combination of the AmpliGrid system, with the AmpliSpeed slide cycler reduces the
amplification reaction time by up to 50% compared to alternative procedures using microplates.
In addition to manual sample application, the AmpliGrid platform is compatible with any kind of
automated liquid handling system making it the platform of choice for high throughput screening
(HTS) applications.
With the AmpliGrid /AmpliSpeed system, Advalytix is setting a new standard for low volume and
single cell assays. When we say single cell, we mean just ONE single cell.
AMPLIGRID AG480F
3
DNA-FREE MICROLITER
REACTION SLIDE
AmpliGrid AG480F
Are you tired of pipetting tiny volumes into microplates? The new AmpliGrid slide
can do the precision work for you!
Pipetting small volumes in microplates often leads to droplets at the sides or the
rims of the wells. Advalytix uses advanced semiconductor lithography to structure
the surface of the AmpliGrid slides in order to allow the specific positioning of one
single microliter (1 µL) on each of the 48 discrete reaction sites. Due to the hydrophilic
nature of the reaction site, all aqueous reagents will be self-centered to the ideal
position, even if not pipetted accurately, making sample handling easy and precise.
AmpliGrid microliter reaction slide.
1 µL reaction volumes can now be your ‘standard’, reducing your reagent and
enzyme consumption by up to 25 times. Furthermore, reducing the reaction volume
also increases the sensitivity and reaction efficiency by a factor of 10, at the same
time reducing the required reaction time.
Pipetting errors will be a matter of the past as the flat surface provides precise optical
control of sample deposition. Due to the high quality 2D-surface any kind of optical
control can be implemented to the AmpliGrid system.
AmpliGrid slides are as easy to use for high throughput procedures, as they are for
low sample numbers when working manually since the technology is easy to integrate
into most of the standard automated liquid handling systems for high throughput
screening (HTS) applications.
Close-up AmpliGrid AG480F.
To enable the full potential of the AmpliGrid system Advalytix will be offering optimised
kits for amplification and reverse transcription. These user-friendly kits are especially
designed for the advanced, low-volume reaction geometry of the AmpliGrid slide.
AmpliGrid Specifications
Size
25 mm x 75.6 mm (± 0.2 mm)
Material
ultra-flat glass microscope slide
Number of reaction sites
48
Diameter reaction sites (hydrophilic area)
1.6 mm (± 0.1 mm)
Diameter reaction sites (hydrophobic area)
3.0 mm (± 0.0 mm)
Features
inactivated, DNA-free
Volume aqueous phase
1 µL (± 10%)
Volume sealing solution
4.8 - 5.0 µL
Pitch reaction sites
4.5 mm (± 0.0 mm)
Manual processing
yes
HTS compatibility
yes
Production facility
class 100 clean room
Cross-section AmpliGrid AG480F.
Optical quality control made easy ...
4
AMPLISPEED ASC100D/200D/400D
ADVANCED TEMPERATURE
MANAGEMENT FOR GLASS SLIDES
ASC100D/200D/400D
The AmpliSpeed slide cycler is an ultra-compact instrument specifically designed for performing
thermal cycling reactions on glass slides. An optimal thermal coupling with the glass slides is
guaranteed using a superior heat transfer technology based on highly-planar silicon wafers. This
unique system generates extremely homogeneous surface temperatures and effective heating and
cooling rates of up to 3°C/second. This can definitely be classified as HIGH-SPEED.
5
AMPLISPEED ASC100D/200D/400D
THERMAL CYCLING: THE NEXT
LEVEL
Advanced technology for excellent ease-of-use and
efficiency
The innovative and intuitive graphical user interface is available either directly on
the slide cycler for benchtop programing, or on a PC for more ambitious use.
With the advanced Ethernet compatibility, the system is easily integrated to your
daily workflow: so while you are working on your PC or enjoying a cup of coffee,
the AmpliSpeed system will send you a reminder when the thermal cycling is
finished. Individual programmes can be stored and transferred using a standard
SD memory card, providing a very flexible memory option for standalone units.
With the AmpliSpeed /AmpliGrid system, Advalytix has defined a new standard
for single cell research and diagnostic applications.
Instruments are available as one, two and four slide units, with each position designed
to run completely independent temperature profiles.
AmpliSpeed slide cycler.
For easy integration into a fully automated liquid handling system, up to 20 units
can be added. All these remarkable features are obtained with whisper-quiet
operation due to the absence of peltier cooling fans.
To achieve maximum accuracy during the amplification processes, an on-slide
temperature calibration feature is available for integration with the AmpliSpeed
system.
AmpliSpeed Specifications
Temperature
Temperature range
ambient +15°C to 99°C
Temperature accuracy
± 0.2°C at 72°C
Temperature homogeneity
± 0.2°C after 20 seconds at 72°C
Effective heating rate
3°C/second
Effective cooling rate
3°C/second
Integration in liquid handling platform.
Software
Temperature ramping
0.1 - 3°C/second
Temperature increments
0.1 - 1°C/cycle
Time increments
0.1 - 60 second/cycle
Memory
SD-card
GLP reports
yes
Instrument
Line Voltage
85 - 230 Volt
Frequency
50 - 60 Hz
Power consumption (ASC100D/200D/400D)
100 Watt maximum
Display
640x480 colour VGA
Auto restart
yes
Dimensions [mm] (wxdxh) ASC100D/200D/400D 126x235x112 /126x295x112 /126x486x112
Weight (ASC100D/200D/400D)
2.0 kg / 2.9 kg /4.7 kg
Operation conditions
+10°C to +30°C
Graphical user interface.
6
DOWNSTREAM APPLICATIONS
APPLICATION EXAMPLES
A
Multiple options for single cells
Genetic typing of single cells
Improved genetic profile.
A Identity testing using human DNA is a growing field in molecular biology. Several
commercial kits and products based on short tandem repeat (STR) analysis of nucleic
acids are available. Using the AmpliGrid technology it is now possible to precisely
apply these kits even for one single cell. Systematic studies to improve sensitivity,
characterise drop-outs, analyse cells in a population one by one as well as many
more approaches have shown excellent new ways for the exact typing of single cells
with this technology.
Single cell sequencing
B
B Sequencing of PCR products generated from a single cell is a challenge in
basic research, immunology, forensics, population genetics as well as other fields
of biology and medicine. With the AmpliGrid, amplification and sequencing from
a defined single cell can be integrated in one workflow. The reaction takes place
exactly where the cell has been deposited. All steps are ‘on-chip’ greatly reducing
the contamination risk and the generation of sequencing artefacts.
Balanced sequencing results.
Copy number variations in single cells
C The analysis of the presence or absence of complete chromosomes as well as
fragments, is fundamental to genetic diagnoses. In many cases, there are only a
limited number of cells available or the genetic answer refers to single cells only
(within mixed populations). Advalytix has developed protocols with significantly
reduced complexity for whole genome amplifications. Applying these methods to
single cells, followed by the redundant analysis of many genetic loci, makes the
detection of the presence/absence of chromosomes and chromatids highly accurate.
C
Combined immunophenotyping and single cell expression
analysis
Analysis of copy number variations.
D
Fluorescence Activated Cell Sorter.
D Single cell RT-PCR is one of the latest techniques used for advanced expression
analysis (see www.gene-quantification.de). From a functional point of view, it is a
great advantage to sort single cells after immunophenotyping, using a fluorescence
activated cell sorter (FACS). Combining the information gathered from a specific
fluorescent marker that reflects a defined function, with this kind of genetic analysis,
is a key breakthrough for the emerging field of ‘cellomics’. The AmpliGrid is the only
amplification platform that can be easily integrated with any cell sorter, allowing high
throughput (and consequently highly redundant) genetic analysis of single cells.
ORDER INFORMATION
7
Order Information
Ref.-No
Product
Description
OAX04503
AmpliGrid AG480F
DNA-free microliter reaction slide (5 pcs./pck.), 48 reaction sites each, incl. 5x500 µL sealing solution
OAX04504
AmpliGrid AG480F
DNA-free microliter reaction slide (5 pcs./pck.), 48 reaction sites each, incl. 1x4 mL sealing solution
OAX04505
AmpliGrid AG480F
DNA-free microliter reaction slide (25 pcs./pck.), 48 reaction sites each, incl. 25x500 µL sealing solution
OAX04506
AmpliGrid AG480F
DNA-free microliter reaction slide (25 pcs./pck.), 48 reaction sites each, incl. 4x4 mL sealing solution
OAX04509
AmpliGrid AG480E
Epoxy coated microliter reaction slide (5 pcs./pck.), 48 reaction sites each, incl. 5x500 µL sealing solution
OAX04510
AmpliGrid AG480E
Epoxy coated microliter reaction slide (5 pcs./pck.), 48 reaction sites each, incl. 1x4 mL sealing solution
OAX04511
AmpliGrid AG480E
Epoxy coated microliter reaction slide (25 pcs./pck.), 48 reaction sites each, incl. 25x500 µL sealing solution
OAX04512
AmpliGrid AG480E
Epoxy coated microliter reaction slide (25 pcs./pck.), 48 reaction sites each, incl. 4x4 mL sealing solution
OAX04513
AmpliGrid DemoKit
Demo Kit DNA-Amplification AG480F
OAX04101
AmpliSpeed ASC100D
AmpliSpeed one bay slide cycler with display unit
OAX04102
AmpliSpeed ASC200D
AmpliSpeed two bay slide cycler with display unit
OAX04103
AmpliSpeed ASC400D
AmpliSpeed four bay slide cycler with display unit
OAX04111
AmpliSpeed ASC100E
one bay extension set for AmpliSpeed systems
OAX04112
AmpliSpeed ASC200E
two bay extension set for AmpliSpeed systems
OAX04113
AmpliSpeed ASC300E
three bay extension set for AmpliSpeed systems
Art. code: AmpliGrid/AmpliSpeed/02-07/O20071001
headquarter
Advalytix AG
Eugen-Sänger-Ring 4, 85649 Brunnthal, Germany
Tel: +49 (0)89 62 83 66 - 0, Fax: +49 (0)89 62 83 66 -10
E-Mail: [email protected]
www.advalytix.de
US office
Frank Feist
58 Elsinore St. Concord, MA 01742, USA
Tel: +1 (978) 405-2533, Fax: +1 (978) 405 2534
Mobile: +1 (978) 979-7899
E-Mail: [email protected]
Advalytix AG is a group company of
Olympus Life and Material Science Europa GmbH