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1/27/2013 10:09 PM
Northern Blot
http://www.molecularstation.com/rna/northern-blot/
Protein Microarray Chips PCR Real-Time PCR RNAi and SiRNA Stem Cell Biology Transfection Western Blot
hybridization probes may be DNA or RNA in northern blotting. A variant of the procedure known as the reverse northern blot was occasionally (although, infrequently) used. In this procedure, the substrate nucleic acid (that is affixed to the membrane) is a collection of isolated DNA fragments, and the probe is RNA extracted from a tissue and radioactively labelled. The use of DNA microarrays that have come into widespread use in the late 1990s and early 2000s is more akin to the reverse procedure, in that they involve the use of isolated DNA fragments affixed to a substrate, and hybridization with a probe made from cellular RNA. Thus the reverse procedure, though originally uncommon, enabled the one-at-a-time study of gene expression using northern analysis to evolve into gene expression profiling, in which many (possibly all) of the genes in an organism may have their expression monitored
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1/27/2013 10:09 PM
Northern Blot
http://www.molecularstation.com/rna/northern-blot/
6. Detection
* *Right before using add 1000ug denatured RNAse free CT DNA (heat to 95o for 3 min to denature) 5000ug yeast RNA
WASH: 2X SSPE + 0.1% SDS (500 mls) 50 mls 20X 5 mls 10% SDS= 0.1% SDS
RNA Gel
After isolating RNA, the RNA must be separated out on a gel (usually agarose). see RNA gels
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1/27/2013 10:09 PM
Northern Blot
http://www.molecularstation.com/rna/northern-blot/
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1/27/2013 10:09 PM
Northern Blot
http://www.molecularstation.com/rna/northern-blot/
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1/27/2013 10:09 PM