title: RNA mapping protocols: Northern blot and amplification of cDNA ends.
abstract: Mapping of transcribed mRNA and protein coding sequences is the initial step in functional characterization of genomic sequences. There are several techniques to identify transcribed sequences; however, Northern blot analysis remains a standard method for detection and quantification of mRNA levels despite the relatively recent availability of other methods, such as RT-PCR (reverse transcriptase polymerase chain reaction), which are also sensitive, accurate, cost-effective, and simple to use. Northern blot analysis provides a direct relative comparison of message abundance between samples on a single membrane. It is also the preferred method for determining transcript size and detecting alternatively spliced isoforms. Other methods utilizing RT-PCR allow detection and cloning of isolated complementary DNA (cDNA) domains and ends from cellular RNA or commercially available cDNA libraries.
authors.affiliation: Diabetes, Cardiovascular, and Metabolic Diseases Division, Translational Genomics Research Institute, Phoenix, AZ, USA.
authors.forename: Maria Lucrecia
authors.initials: ML
authors.lastname: Alvarez
authors.forename: Mahtab
authors.initials: M
authors.lastname: Nourbakhsh
mesh_terms: D015152:Blotting, Northern; D018076:DNA, Complementary / Q000235:genetics*; D055106:Genome-Wide Association Study / Q000379:methods*; D006801:Humans; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger