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Northern Analysis: The Basics

ULTRAhyb-Oligo has been specifically formulated for use with oligonucleotide probes in Northern analysis. ULTRAhyb-OS is available for use with infrared probes detected by the LI-COR Odyssey Infrared Imaging System.


Washing

After hybridization, unhybridized probe is removed by washing in several changes of buffer. Low stringency washes (e.g., with 2X SSC or SSPE) remove the hybridization solution and unhybridized probe. High stringency washes (e.g., with 0.1X SSC or SSPE) remove partially hybridized molecules. Certified RNase-free low and high stringency wash buffers are included in the NorthernMax Kits, and are also available separately.


Detection

If a radiolabeled probe was used, the blot can be wrapped in plastic wrap to keep it from drying out and then immediately exposed to film for autoradiography. If a nonisotopic probe was used, the blot must be treated with nonisotopic detection reagents prior to film exposure. The BrightStar BioDetect Nonisotopic Detection Kit provides all the reagents and materials necessary for detection of biotinylated
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