公开(公告)号：US20110015096A1

公开(公告)日：2011-01-20

申请号：US12610749

申请日：2009-11-02

Applicant: KUO PING CHIU

Inventor： KUO PING CHIU

IPC: C40B40/06 ,  C07H21/00 ,  C40B50/00

CPC classification number: C40B50/06 ,  C12N15/1093 ,  C12N15/66 ,  C40B40/06

Abstract: Multiplex barcoded Paired-End Ditag (mbPED) library construction for ultra high throughput sequencing is disclosed. The mbPED library comprises multiple types of barcoded Paired-End Ditag (bPED) nucleic acid fragment constructs, each of which comprises a unique barcoded adaptor, a first tag, and a second tag linked to the first tag via the barcoded adaptor. The two tags are the 5′- and 3′-ends of a nucleic acid molecule from which they originate. The barcoded adaptor comprises a barcode, a first polynucleotide sequence comprising a first restriction enzyme (RE) recognition site, and a second polynucleotide sequence comprising a second RE recognition site and covalently linked to the first polynucleotide sequence via the barcode. The two REs lead to cleavage of a nucleic acid at a defined distance from their recognition sites. The length of the adaptor is set so that the bPED nucleic acid fragment fits one-step sequencing.

Abstract translation: 公开了用于超高通量测序的多路复用条形码配对结束Ditag（mbPED）库构建。 mbPED库包含多种类型的条形码配对结束分析（bPED）核酸片段构建体，每个构建体包含独特的条形码适配器，第一标签和通过条形码适配器链接到第一标签的第二标签。 两个标签是来源于其的核酸分子的5'和3'末端。 条形适配器包括条形码，包含第一限制酶（RE）识别位点的第一多核苷酸序列和包含第二RE识别位点的第二多核苷酸序列，并经由条形码共价连接到第一多核苷酸序列。 两个RE导致在与其识别位点限定距离处的核酸裂解。 适配器的长度设定为使得bPED核酸片段适合一步测序。