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METHODS AND COMPOSITIONS FOR cDNA SYNTHESIS AND SINGLE-CELL TRANSCRIPTOME PROFILING USING TEMPLATE SWITCHING REACTION

机译:利用模板交换反应进行cDNA合成和单细胞转录谱分析的方法和组成

摘要

This application discloses methods for cDNA synthesis with improved reverse transcription, template switching and preamplification to increase both yield and average length of cDNA libraries generated from individual cells. The new methods include exchanging a single nucleoside residue for a locked nucleic acid (LNA) at the TSO 3′ end, using a methyl group donor, and/or a MgCl2 concentration higher than conventionally used. Single-cell transcriptome analyses incorporating these differences have full-length coverage, improved sensitivity and accuracy, have less bias and are more amendable to cost-effective automation. The invention also provides cDNA molecules comprising a locked nucleic acid at the 3′-end, compositions and cDNA libraries comprising these cDNA molecules, and methods for single-cell transcriptome profiling.
机译:该申请公开了具有改进的逆转录,模板转换和预扩增以增加从单个细胞产生的cDNA文库的产量和平均长度的cDNA合成方法。新方法包括使用甲基供体和/或高于常规使用的MgCl 2 浓度,在TSO 3'末端将单个核苷残基交换为锁定核酸(LNA)。融合了这些差异的单细胞转录组分析具有全长覆盖范围,更高的灵敏度和准确性,更小的偏倚以及更符合成本效益的自动化方法。本发明还提供了在3'端包含锁定核酸的cDNA分子,包含这些cDNA分子的组合物和cDNA文库,以及用于单细胞转录组谱分析的方法。

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