醛硅烷涂层、DNA芯片

Aldehydesilane coating

NEXTERION^® Slide AL

NEXTERION^® Slide AL offers a good alternative to NEXTERION^® Slide E (Epoxysilane slides) when an efficient covalent, and directed binding of amino-modified bio-molecules is required. NEXTERION^® Slide AL is coated with an “active” slide chemistry coating that chemically binds bio-molecules to provide a high binding capacity, and minimize non-specific binding. The uniform surface features aldehyde groups that readily react with primary amines. Both 5’ or 3’ amine modified PCR products, and oligonucleotides work well with this surface chemistry. The covalent bond formed at the terminus of the nucleic acid offers both stability, and maximal base pairing opportunity. The covalent binding diminishes sample loss during the course of experiments, and permits more harsh wash steps, which reduce background, and allow for greater sensitivity. Additional immobilization steps, such as baking or UV cross-linking, are not required for immobilization. Furthermore, peptides, proteins (such as antibodies), cells, and tissues can be immobilized on NEXTERION^® Slide AL via random binding to amine sites on the probes. The slides are easy to use, and are compatible with all commercially available arraying and scanning instruments.

Overview
 

Suitable probe types

• Amine-PCR products and BACs

• Amino-modified oligonucleotides

• Small protein fragments such as peptides

• Cells and tissues

Immbilization chemistry

The DNA product is spotted onto the aldehydesilane slide. The primary amino linkers (NH₂) on the DNA attack the aldehyde groups to form covalent bonds (Schiff’s base). The attachment is stabilized by a dehydration reaction. To minimize fluorescent background, unreacted aldehyde groups are reduced to non-reactive primary alcohols by treatment with sodium borohydride. In addition, this step reduces the double bond between the probe and surface, producing an irreversible covalent immobilization. Additional steps for immobilization, such as baking, or UV cross-linking, are not required.

 

Product details

Highly reproducible coating

NEXTERION^® Slide AL are produced using an innovative process developed by SCHOTT to produce a consistent and reproducible coating. All slides are individually examined for physical defects and the presence of particles before and after coating. The density of the aldehyde groups on the coating remain constant over the entire surface of the slides, and has been optimized to provide the optimal binding capacity. The surface hydrophobicity is tightly controlled to optimize the performance with both contact and non-contact printers.

High intra-slide reproducibility of NEXTERION^® Slide AL (20 k array of BSA labelled with Atto 547)

Packaging and storage

NEXTERION^® Slide AL are packaged in chemically stable plastic boxes and sealed in an inert atmosphere. The slides are ready-to-use from the box, and are stable for 9 months in the sealed packaging when stored at room temperature.

Format

The slides are available in packs of 25-slides with optional code 128 barcodes enabling automated sample tracking. The aldehydesilane coating is also available in 16-well slide, 48-well slide and 96-well 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	NEXTERION® Spot + detergents like cetyltrimethylammoniumbromid, Triton X-100, sarcosyl, Tween or SDS with a final concentration between 0.005 and 0.05 % (to increase the spot size)	Recommended Spotting Concentrations: PCR products: 0.1 – 1 µg/µL Oligonucleotides: 10 – 20 µM Peptides: 100 – 500 µg/mL Recommended Spotting Conditions: Constant relative humidity 40 – 50% at 20 – 24°C (68 to 75°F). Spotting solutions may be used with, or without protein-stabilizing agents
		3x SSC
		3x SSC + 1.5 M betaine (low evaporation buffer for long spotting runs)
Chemical deactivation	-	1.0 g NaBH4 in 300 mL PBS and 100 mL ethanol
Hybridization	NEXTERION®Hyb (formamide free) NEXTERION®Oligo Hyb (with formamide)	3 – 5x SSC + 0.1 % SDS	Add competitor DNA if appropriate

 

NEXTERION^® Slide AL Evaluation Kit
 

NEXTERION^® Slide AL is easy to use, but as this type of slide chemistry may be less familiar to some researchers, a dedicated NEXTERION^® Slide AL Evaluation Kit is available. The kit consists of slides with test oligonucleotides and optimized reagents, allowing new users to carry out a test print and hybridization.

Process step	SCHOTT product	Quantity
Spotting	NEXTERION® Slide AL	10 slides
	NEXTERION® Spot	10 mL
Hybridization	NEXTERION® Hyb	10 mL
	Oligo test probe	2 nmol
	Oligo test target	3 pmol