title: Atomic force microscope observation of branching in single transcript molecules derived from human cardiac muscle.
abstract: We have used an atomic force microscope to examine a clinically derived sample of single-molecule gene transcripts, in the form of double-stranded cDNA, (c: complementary) obtained from human cardiac muscle without the use of polymerase chain reaction (PCR) amplification. We observed a log-normal distribution of transcript sizes, with most molecules being in the range of 0.4-7.0 kilobase pairs (kb) or 130-2300 nm in contour length, in accordance with the expected distribution of mRNA (m: messenger) sizes in mammalian cells. We observed novel branching structures not previously known to exist in cDNA, and which could have profound negative effects on traditional analysis of cDNA samples through cloning, PCR and DNA sequencing.
authors.affiliation: Department of Chemistry and Biochemistry, UCLA, 607 Charles Young Drive East, Los Angeles, CA 90095, USA. California NanoSystems Institute (CNSI), 570 Westwood Plaza, Los Angeles, CA 90095, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: Reed
authors.forename: Carlin
authors.initials: C
authors.lastname: Hsueh
authors.forename: Bud
authors.initials: B
authors.lastname: Mishra
authors.forename: James K
authors.initials: JK
authors.lastname: Gimzewski