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Atomic force microscope observation of branching in single transcript molecules derived from human cardiac muscle

Jason Reed1,2,4, Carlin Hsueh1, Bud Mishra3 and James K Gimzewski1,2,4

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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.


PACS

87.64.Dz Scanning tunneling and atomic force microscopy

87.19.Ff Muscles

87.14.G- Nucleic acids

36.20.Fz Constitution (chains and sequences)

87.19.Hh Cardiac dynamics

87.15.B- Structure of biomolecules

Subjects

Soft matter, liquids and polymers

Atomic and molecular physics

Medical physics

Biological physics

Dates

Issue 38 (24 September 2008)

Received 21 March 2008, in final form 29 May 2008

Published 12 August 2008



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