Abstract:
:Subtractive hybridization has been widely used for the identification of differentially expressed genes. Here we describe a simple, sensitive strategy of subtractive hybridization that involves binding the driver poly(A)+ RNA pool to paramagnetic Dynabeads Oligo (dT)25. After hybridization with target cDNA, the molecules common to both pools are removed. The subtracted cDNA is then amplified with PCR and used for library screening. Using this method, we have identified four cDNA clones that represent developmentally regulated transcripts in the central nervous system of the tobacco hornworm Manduca sexta. All four transcripts are of low abundance, comprising only 0.001%-0.5% of the poly(A)+ RNA pool.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Mészáros M,Morton DBdoi
10.2144/19962003413subject
Has Abstractpub_date
1996-03-01 00:00:00pages
413-9issue
3eissn
0736-6205issn
1940-9818journal_volume
20pub_type
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