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 storageNEXTERION® 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.FormatThe 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 |