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    Whole transcriptome sequencing
    9.
    发明授权
    Whole transcriptome sequencing 有权
    全转录组测序

    公开(公告)号：US09441264B2

    公开(公告)日：2016-09-13

    申请号：US14074958

    申请日：2013-11-08

    申请人： Genisphere, LLC

    发明人： Robert C. Getts ,  James Kadushin

    IPC分类号： C12Q1/68 ,  C12N15/10

    CPC分类号： C12Q1/6806 ,  C12N15/1096

    摘要： Methods for preparing strand-specific sequencing libraries of oligonucleotides using an RNA polymerase promoter to re-transcribe antisense cDNA which has been reverse transcribed from mRNA are provided. The transcription step linearly amplifies sRNA prior to production of double-stranded cDNA to be sequenced and may be sufficient to eliminate the conventional PCR amplification step prior to sequencing. The methods incorporate anchor sequences, amplification sequences and other sequences required for a particular sequencing system or reaction by hybridization and extension of primers, and transcription of RNA, rather than ligation, thus reducing the number of steps and the time required for sample preparation for sequencing of RNA. Use of primer hybridization and transcription reactions in the methods also results in a library that exhibits reduced 3′ sequence bias.

    摘要翻译： 提供了使用RNA聚合酶启动子制备寡核苷酸的链特异性测序文库以重新转录已经从mRNA逆转录的反义cDNA的方法。 转录步骤在产生待测序的双链cDNA之前线性扩增sRNA，并且可能足以在测序之前消除常规的PCR扩增步骤。 该方法包含锚定序列，扩增序列和特定测序系统所需的其他序列或通过引物杂交和扩增反应以及RNA的转录而不是连接，从而减少样品制备测序所需的步骤数和时间 的RNA。 在该方法中使用引物杂交和转录反应也导致显示减少的3'序列偏差的文库。

    Whole Transcriptome Sequencing
    10.
    发明申请
    Whole Transcriptome Sequencing 有权
    全转录片段测序

    公开(公告)号：US20140113333A1

    公开(公告)日：2014-04-24

    申请号：US14074958

    申请日：2013-11-08

    申请人： Genisphere, LLC

    发明人： Robert C. Getts ,  James Kadushin

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6806 ,  C12N15/1096

    摘要： Methods for preparing strand-specific sequencing libraries of oligonucleotides using an RNA polymerase promoter to re-transcribe antisense cDNA which has been reverse transcribed from mRNA are provided. The transcription step linearly amplifies sRNA prior to production of double-stranded cDNA to be sequenced and may be sufficient to eliminate the conventional PCR amplification step prior to sequencing. The methods incorporate anchor sequences, amplification sequences and other sequences required for a particular sequencing system or reaction by hybridization and extension of primers, and transcription of RNA, rather than ligation, thus reducing the number of steps and the time required for sample preparation for sequencing of RNA. Use of primer hybridization and transcription reactions in the methods also results in a library that exhibits reduced 3′ sequence bias.

    摘要翻译： 提供了使用RNA聚合酶启动子制备寡核苷酸的链特异性测序文库以重新转录已经从mRNA逆转录的反义cDNA的方法。 转录步骤在产生待测序的双链cDNA之前线性扩增sRNA，并且可能足以在测序之前消除常规的PCR扩增步骤。 该方法包含锚定序列，扩增序列和特定测序系统所需的其他序列或通过引物杂交和扩增反应以及RNA的转录而不是连接，从而减少样品制备测序所需的步骤数和时间 的RNA。 在该方法中使用引物杂交和转录反应也导致显示减少的3'序列偏差的文库。