公开(公告)号：US20150337298A1

公开(公告)日：2015-11-26

申请号：US14719149

申请日：2015-05-21

Applicant: FLUIDIGM CORPORATION

Inventor： Lei Xi ,  Xiaohui Wang ,  Marc Unger ,  David Ruff

IPC: C12N15/10 ,  C12Q1/68

CPC classification number: C12N15/1082 ,  C12N15/1065 ,  C12Q1/6806 ,  C12Q2525/155 ,  C12Q2525/301 ,  C12Q2535/122

Abstract: In certain embodiments, the present invention provides a way of “digitally” marking different the alleles of different chromosomes by using a transposase to insert differently barcoded transposons into genomic DNA before further analysis. According to this method, each allele becomes marked with a unique pattern of transposon barcodes. Because each unique pattern of transposon barcodes identifies a particular allele, the method facilitates determinations of ploidy and copy number variation, improves the ability to discriminate among homozygotes, heterozygotes, and patterns arising from sequencing errors, and allows loci separated by uninformative stretches of DNA to be identified as linked loci, thereby facilitating haplotype determinations. Also provided is a novel artificial transposon end that includes a barcode sequence in two or more positions that are not essential for transposition.

Abstract translation: 在某些实施方案中，本发明提供了一种通过使用转座酶在进一步分析之前将不同条形码转座子插入基因组DNA中来“不同”地标记不同染色体等位基因的方式。 根据这种方法，每个等位基因都被标记为转座子条形码的独特模式。 因为转座子条形码的每个独特模式识别特定的等位基因，所以该方法有助于确定倍性和拷贝数变异，提高了鉴别纯合子，杂合子和测序错误引起的模式的能力，并且允许通过DNA的非信息延伸分离的位点 被确定为相关基因座，从而促进单倍型测定。 还提供了一种新颖的人工转座子末端，其包括对于转座不是必需的两个或更多个位置的条形码序列。