Aminosilane coating


NEXTERION® Slide A+




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. With it's NEXTERION® Slide A+ SCHOTT is committed to providing the best aminosilane microarray slides on the market.


Overview

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 A+ None 1064875 25
Laser 1064877 25
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
Typical applications
• ArrayCGH
• Transcriptional profiling
• SNP genotyping
• Splice variant detection
• DNA methylation profiling
Suitable probe types
• BACS or PACs
• Oligonucleotides ≥ 40 mers
• Small protein fragments such as peptides
• cDNA


Introduction

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.
Linker length of NEXTERION aminosilane coated microarray slides
Comparison of aminosilane structures
The aminosilane slide offered by SCHOTT is based on very short chain aminosilanes that demonstrate high signal intensities, and exceptionally low background signals, when compared with other commercially available aminosilane slides.



Immobilization chemistry

Signal-to-background ratios of aminosilane coated microarray slides
Signal-to-background ratios compared to market leading aminosilane slide
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.
linking probes to aminosilane coated microarray slides


Product details

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 coating is 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 coating remains 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.
NEXTERION® Slide A+ is 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.


Packaging and storage
NEXTERION® Slide 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 9 months when stored at room temperature in the
sealed packaging.

Format
NEXTERION® Slide A+ are available in packs of 25 slides with optional code 128 barcodes enabling automated sample tracking. The NEXTERION® A+ coating is also available in multi-well slide and microplate formats. For further information, refer to the section on “Multi-well formats”.

Compatible reagents

Protocol
step
Recommended NEXTERION® products Alternatives Additional information
Spotting NEXTERION® Spot
(1066029)

NEXTERION® Spot A HD
(Composition can be provided)

Recommended Spotting Concentrations:
Oligonucleotides 2 – 20 µM
PCR Products 0.5 – 1 mg/mL
Recommended Spotting Conditions:
Constant 40 – 50 % relative humidity at 20 to 25 °C
25 – 50 % DMSO
3x SSC 
3x SSC containing

1.5 M betaine 
Blocking   NEXTERION® Block A Kit
(Composition can be provided)
 
Hybridization NEXTERION® Hyb
(1066075)
NEXTERION® Oligo Hyb
(Composition can be provided)
3 – 5x SSC + 0.1 % SDS
Add competitor DNA if appropriate

Protocols

Protocol revision
Date: April 2009
Protocol version: 1.4
Revision made: minor layout changes, new telephone and fax numbers
Revision reason: correction contact information



Material Safety Datasheets

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