The Ambion® NorthernMax®-Gly Kit is an alternative to formaldehyde-based systems for northern analysis and uses a glyoxal/DMSO Loading Solution for sample denaturation. Sufficient reagents are provided for processing 1000 cm2 membrane.
• Glyoxal gels can be prepared at the lab –bench—no noxious reagents
• Increase sensitivity up to 100x over standard hybridization protocols
• Reduce hybridization to just 2 hours for many messages
• Complete kit contains ULTRAhyb® Hybridization Buffer and gel and wash reagents for 10–20 gels
• Compatible with RNA, DNA, or oligonucleotide –probes—labeled isotopically or nonisotopically
Use of glyoxal/DMSO instead of formaldehyde for sample preparation avoids the need to pour and run gels in a fume hood, and eliminates the safety issues associated with use of formaldehyde. Also, RNA samples denatured with glyoxal may show sharper bands on northern blots compared to samples denatured and run in the presence of formaldehyde. The streamlined NorthernMax®-Gly procedure is more convenient and faster than the standard glyoxal gel protocol. A single reagent is used for sample denaturation and loading, the incubation time for sample denaturation has been reduced, and no recirculation of buffer is required during electrophoresis. The volume ratio of glyoxal denaturation solution to sample RNA is lower than in other published protocols, so that sample precipitation prior to gel loading is usually not required. The NorthernMax®-Gly Kit can be used with either radiolabeled or nonisotopically labeled RNA or DNA probes. Each NorthernMax®-Gly Kit provides a complete system. Positively charged nylon membranes must be purchased separately (see Accessory Products). An RNA sample and template for probe synthesis are also provided as controls for each kit.
Accessory Products:
The BrightStar® -Plus Positively Charged Nylon Membranes (SKU# AM10100, AM10102, or AM10104) are optimized for use with the NorthernMax®-Gly Kit.
Contents and Storage
Positive Control RNA, pTRI-GAPDH Mouse, DECAtemplate §-actin-Mouse, and Glyoxal Load Dye should be stored at –20°C. ULTRAhyb® Buffer and 10X Gel Prep⁄Running Buffer should be stored at 4°C. Agarose-LE™, Transfer Buffer, Low Stringency Wash Solution #1, mL High Stringency Wash Solution #2, and RNaseZap® should be stored at room temperature. Nuclease-free water may be stored at any temperature.
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