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

机译:原子力显微镜观察人心肌单个转录本分子中的分支

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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.
机译:我们已经使用原子力显微镜检查了从人心肌中获取的单分子基因转录物的临床衍生样品,其形式为双链cDNA(c:互补),而未使用聚合酶链反应(PCR)扩增。我们观察到转录本大小的对数正态分布,根据mRNA的预期分布(m:信使),大多数分子的轮廓长度在0.4-7.0千碱基对(kb)或130-2300 nm范围内在哺乳动物细胞中。我们观察到了以前未知的cDNA中不存在的新型分支结构,并且可能通过克隆,PCR和DNA测序对cDNA样品的传统分析产生深远的负面影响。

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