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SCHOTT North America
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Aminosilane coating
Nexterion® Slide A+ and Slide AStar
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Aminosilane coated slides remain the most popular choice for printing PCR
products, and long oligonucleotides, despite the emergence of innovative
three-dimensional microarray surfaces, and other “active” surface chemistries
such as epoxysilane. SCHOTT is committed to providing the best aminosilane
microarray slides on the market, and offers two types of aminosilane slides,
Nexterion® Slide A+, and Slide AStar.
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| Type of
coating |
Immobilization
method |
Typical
probes |
Ordering information |
| Nexterion® product |
Barcode
option |
Item
number |
Slides
per pack |
Aminosilane
2-D surface |
Ionic interaction followed
by cross-linking via an
additional UV or baking step |
• Long oligonucleotides
• PCR products
• BACs |
Slide AStar |
None |
1177881 |
25 |
| Laser |
1177882 |
25 |
| Slide A+ |
None |
1064875 |
25 |
| Laser |
1064877 |
25 |
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| Key product features |
| • Printed slides have a long shelf life |
| • Compatible with a wide range of spotting buffers |
| • Coatings with uniform aminosilane density |
| • Regular spot uniformity and morphology |
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| Typical applications |
| • ArrayCGH |
| • Transcriptional profiling |
| • SNP genotyping |
| • Splice variant detection |
| • DNA methylation profiling |
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| Suitable probe types |
| • BACS or PACs |
| • Oligonucleotides ≥ 40 mers |
| • Small protein fragments such as peptides |
| • cDNA |
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Introduction
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Aminosilane surfaces provide available
amine groups for initial ionic attachment of the negatively charged phosphate
groups in the DNA backbone. Conventional wisdom is that longer aminosilane
molecules with multiple amino groups produce higher signal-to-background
ratios, as their higher positive charge will bind more DNA to the slide surface.
However, recent research at SCHOTT has shown that the situation is actually
more complicated than this, and that while surfaces with a higher charge do
bind more DNA, they are also more difficult to block effectively, resulting in
significantly higher non-specific binding. The higher levels of non-specific
binding have a negative effect leading to poorer signal-to-background ratios.
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Comparison of aminosilane structures
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The two aminosilane slides now offered by SCHOTT are based on very short
chain aminosilanes that demonstrate high signal intensities, and exceptionally
low background signals, when compared with other commercially available
aminosilane slides.
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Signal-to-background ratios
compared to market leading aminosilane slide
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Aminosilane coated slides have a high concentration of primary aminogroups
available at the surface. These groups become protonated and therefore
positively charged when placed in contact with a near-neutral, aqueous
solution. Negatively charged probe molecules, such as DNA, will initially form
multiple ionic interactions with the positively charged amino surface coating.
Additional amino-modifications of the nucleic acids are not required, but such
modifications do not interfere with the immobilization. After spotting, the probes are covalently linked to the slide surface by either heating, or a brief
exposure to ultraviolet (UV) light. Other types of negatively charged biomolecules
may also be coupled to aminosilane surfaces.
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Product details
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Reproducible results
SCHOTT slides are manufactured from a high quality, low intrinsic fluorescence
borosilicate glass. The glass slides are cleaned and coated in a class 100, environmentally
controlled clean room to ensure contamination and artifact free
surfaces. The aminosilane coatings are applied using a unique, and innovative
method developed and optimized by SCHOTT, that allows the production of
large lot sizes with excellent intra-lot, and inter-lot reproducibility. Each slide
lot is tested using both physical and functional quality control checks. The
density of the aminosilane groups in the coatings remain uniform over the
entire surface of the slides, and is optimized to maximise the DNA binding
capacity. The surface hydrophobicity is tightly controlled to optimize the performance
with both contact and non-contact microarray printers. The Slide A+
surface is more hydrophobic than the AStar surface, resulting in smaller spots,
making it ideally suited for printing higher density arrays.
Nexterion® Slide A+, and Slide AStar are compatible with the most commonly
used aminosilane protocols and a wide range of spotting buffers. This makes
it easy to evaluate and switch to the Nexterion® aminosilane slides from competitor
slides.
Selecting the most suitable Nexterion® aminosilane coating
Choosing the best SCHOTT aminosilane slide for the application depends
on a number of factors. One important factor is the buffer used for printing
the arrays. SCHOTT has developed a printing buffer system (Nexterion®
Spot A HD) for printing high-density arrays
of up to 50k spots on the SCHOTT aminosilane slides. The buffer system is
designed to minimize sample evaporation during long print runs.
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Nexterion® Slide A+ |
Nexterion® Slide AStar |
| Compatible print buffers |
Nexterion® Spot A HD
Nexterion® Spot
3x SSC
3x SSC containing 1.5 M betaine
25 – 50 % DMSO
Pronto!™ Universal Spotting Solution |
Nexterion® Spot A HD
Nexterion® Spot
25 – 50 % DMSO
Pronto!™ Universal Spotting Solution |
| Spot diameter |
80 µm
(Nexterion® Spot A HD) |
90 µm
(Nexterion® Spot A HD) |
| High hydrophobicity |
+++ |
++ |
| Suitable for high density applications |
+++ |
++ |
| High signal yield |
+++ |
+++ |
| Low non-specific binding |
++ |
+++ |
| High signal-to-background |
+++ |
++++ |
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Packaging and storage
Nexterion® Slide AStar and A+ are packaged in specially developed compatible
plastic boxes, and sealed under an inert atmosphere, to ensure the substrates
have a long and stable shelf life. The slides are ready-to-use from the box,
and are stable for up to 12 months when stored at room temperature in the
sealed packaging.
Format
Nexterion® Slide AStar and A+ are available in packs of 25-slides with optional
code 128 barcodes enabling automated sample tracking. The AStar and A+
aminosilane coating are also available in 16-well slide and 96-well microplate
formats. For further information refer to the section on “Multi-well formats”.
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Nexterion® Aminosilane Slide
Processing Kit
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SCHOTT Microarray Solutions offers a kit with pre-prepared reagents for the
blocking, hybridization and washing of 25 aminosilane coated slides printed
with oligonucleotides.
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| Process step |
SCHOTT product |
Quantity |
| Blocking |
Nexterion® Block A Kit |
500 mL |
| Hybridization |
Nexterion® Hyb |
10 mL |
| Washing |
Nexterion® Wash A |
1000 mL |
| Nexterion® Wash B |
100 mL |
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Protocol revision
Date: April 2009
Protocol version: 1.3
Revision made: minor layout changes, new telephone and fax numbers
Revision reason: correction contact information
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Protocol revision
Date: April 2009
Protocol version: 1.4
Revision made: minor layout changes, new telephone and fax numbers
Revision reason: correction contact information
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Nexterion® Slide A is available on request
Protocol revision
Date: April 2009
Protocol version: 1.2
Revision made: minor layout changes, new telephone and fax numbers
Revision reason: correction contact information
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Please request the MSDS
by completing the form.
(* mandatory fields)
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Contact
Microarray Solutions
SCHOTT North America Inc.
5530 Shepherdsville Road
Louisville, KY 40228
USA
 | +1 -502-657-4411 |
 | +1 -502-966-4976 |
E-mail
More contacts
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