Arrayit SuperNylon Microarray Substrate Slides (Box of 25) provide atomically smooth glass substrate slides coated with a 150 µm layer of SuperNylon membrane. SuperNylon substrate slides support the standard glass slide format (1 x 25 x 76 mm) and have a corner chamfer to allow unambiguous orientation during use. A time-tested membrane surface for a wide variety of fluorescent and colorimetric DNA and protein microarray applications, provides small printed spots, strong and uniform signals, and low background. Use in conjunction with the colorimetric SpotWare scanner and Arrayit's extensive line of microarray products and services, 25 slides per box.
Microarray Slides - SuperNylon Microarray Substrate Slides for DNA and Protein Microarray Manufacturing with Colorimetric and Fluorescent Detection
Arrayit SuperNylon Microarray Substrate Slides provide a highly innovative nylon membrane surface for researchers seeking the time-tested attributes of a positively charged nylon membranes in a microarray format (1 x 25 x 76 mm). Arrayit SuperNylon can be used to attach DNA, proteins, carbohydrates and all other molecules that bind to nylon in traditional filter assays. The high binding capacity of the 150 µm nylon layer is complimented by a polished atomically smooth glass substrate slide for optimal performance compatibility with all standard glass substrate slide microarray printers and scanners.
Description
- Positively charged membrane-coated glass substrate slide (1 x 25 x 76 mm)
- Corner chamfer for side orientation
- 150 µm positively charged nylon layer
- Binding capacity DNA = 5 µg per square millimeter
- Binding capacity Protein = 2 µg per square millimeter
- Reaction area 25 x 76 mm
Arrayit SuperNylon is compatible with all glass slide microarray scanners including the Arrayit InnoScan Scanners, Arrayit SpotLight Microarray Scanners and the Arrayit SpotWare Colorimetric Scanners.
The following detection methods can be used with this surface:
- Fluorescence
- Colorimetric
- Chemiluminescence
- Radioactivity
Arrayit SuperNylon Substrate Slides are ready to use “right out of the box” without any additional processing or surface preparation. The surface is positively-charged to allow crosslinking of printed DNA and protein molecules to the surface. Arrayit nylon surfaces are fully compatible with a variety of manufacturing technologies including Arrayit contact printing. BlockIt Blocking Buffer should be used for low background and to prevent non-specific binding in complex assays and hybridization reactions. DNA microarrays wash steps can be performed using Wash Buffers A, B and C.
SuperNylon Usage Tips
Microarray Printing:
Proteins should be printed in Protein Printing Buffer. DNA should be printed in Micro Spotting Plus. If the protein is already in a 1X PBS solution, add an equal volume of printing buffer to achieve a final protein concentration between 0.1-2.0 µg/µl. Lyophilized oligonucleotides can be printed at 1-50 µM final concentration with phenol red (0.001%) as the tracking dye to monitor the microarray printing quality. DNA crosslinking should be done with 1,200 J of UV. Allow printed microarray spots do dry before cross linking.
Arrayit SuperNylon contains a 150 µm thick layer of nylon membrane that is positively charged and has a high binding capacity. When using a robotic microarray printer such as the NanoPrint 2, SpotBot 4 or other microarrayers in conjunction with Arrayit contact printing devices, make sure to decrease the Z-axis speed to 2 cm/sec or less to avoid denting the membrane. Because of the hydrophilic nature of this substrate slide, maintain minimal dwell times of of the microarray pins on the surface during printing. Print at 50-80% relative humidity and 20-25°C. To increase the integrity of molecules in the 384-well microplates, use a system like the SpotBot 4 or NanoPrint 2 with source plate cooling set to 10-15°C.
Printed Microarray Storage:
Printed microarrays should be incubated overnight at low humidity to drive the coupling reaction of printed molecules to the nylon membrane. Printed DNA spots must be dry prior to crosslinking to the surface. Microarrays should be stored in slide boxes in a clean, dry and cool location. Printed microarrays can also be stored at 4°C or -20°C for extended storage. Vacuum packing can also be used to enhance printed microarray storage. Make sure microarrays are dry prior to vacuum packing. Store printed microarrays unblocked and un-washed, and proceed to blocking and washing steps just prior to incubation or hybridization.
Arrayit SuperNylon Blocking and Washing:
Prior to performing binding reactions, microarrays printed on Arrayit SuperNylon Substrate Slides should be blocked with BlockIt buffer for 1 hour and then washed 3 times in 1X Protein Microarray Wash Buffer for 5 minutes using a high throughput wash station. The surface should remain hydrated at all times between the blocking, binding and washing reactions. Arrayit SuperNylon should only be allowed to dry after printing and prior to scanning. Arrayit Microarray High Speed Centrifuges (Cat. MHC) can be used for rapid drying.
SuperNylon Incubation and Detection Reactions:
Binding reactions can be 1 hour to overnight. Experiments must not be allowed to dry
out. Fluorescent reactions running for long periods of time should be kept in the dark. Secondary antibody reactions should be run for up to 2 hours and typically not less than 30 minutes. Concentration of detection antibody needs to be optimized empirically by the user. Colorimetric and other enzymatic detection reactions require optimization by the user. Biotin-labeled cRNAs, known as labeled cRNA targets, can be generated using TrueLabeling-AMP Linear RNA Amplification Kit following manufacturer’s protocol (SuperArray Bioscience Corporation). Briefly, total RNA (3 µg/array) can be converted to cDNA at 42°C for 50 minutes. These cDNAs are then in vitro transcribed to cRNAs in the presence of biotin-16-UTP (Roche Molecular Biochemicals, Basel, Switzerland, http://www.roche.com). Biotin-labeled cRNAs can be purified using a RNeasy Mini Kit (Qiagen). The concentration of cRNAs can be measured with a UV spectrophotometer. Microarrays can be hybridized with biotin-labeled targets (5 µg per microarray) at 60°C for 1-4 hours. Filters can be washed with Wash buffers A, B and C at 60°C for 15 minutes each. Chemiluminescent detection steps can be performed by subsequent incubation of the microarrays with alkaline phosphatase–conjugated streptavidin and CDP-Star substrate.
Scanning:
Arrayit SuperNylon Microarray Substrate Slides are compatible with a variety of detection methods. Microarray slides should be scanned dry unless the detection method is chemiluminescent. Slides can be air-dried in the dark or spun dry using an Arrayit Microarray High-Speed Centrifuge (Cat. MHC). The nylon membrane should appear white prior to scanning. When scanning SuperNylon and other white membrane surfaces using laser-photomultiplier tube (PMT) microarray scanners, the default laser/PMT used for glass transparent glass slides may not be optimal for membrane slides. Because of the increased binding capacity of the nylon surface and increased reflection, lower laser power and PMT settings should be used. For results in fluorescent detection, use Arrayit Microarray Scanners.
Recommended Equipment and Reagents
NanoPrint™ 2 Microarrayers
SpotBot® 4 Personal Microarrayers
InnoScan® Microarray Scanners
SpotLight™ 2 Microarray Scanners
Microarray Hybridization Cassettes
High Throughput Wash Stations
Microarray High-Speed Centrifuge
BlockIt™ Blocking Buffer
Microarray Air Jet
Microarray Cleanroom Wipes
PCR Purification Kits
Micro-Total RNA Extraction Kit
MiniAmp mRNA Amplification Kit
Indirect Amino Allyl Fluorescent Labeling Kit
Universal Reference mRNA
Green540 and Red640 Reactive Fluorescent Dyes
Hybridization Buffers