Complete Genomics, Inc.

Mgi Tech Co., Ltd.

Andrei Alexeev

Radoje Drmanac

Brock A Peters

Peter Hong

Hui Fu Jiang

Wenwei Zhang

Yuan Jiang

Xia Zhao

Dong Li

Chunyu Geng

Tong Wang

Yingxin Zhang

Qiaoling Li

Lingyu He

Chao Dong

Rongrong Guo

Evan Hurowitz

Ruoying Chen

Shujin Fu

Yaqiao Li

Xiaoshan Su

Fanzi Wu

Hui Jiang

Jie Ruan

The present invention provides methods and compositions for tagging long fragments of a target nucleic acid for sequencing and analyzing the resulting sequence information in order to reduce errors and perform haplotype phasing, for example.

Multiple tagging of long DNA fragments

The present invention is directed to methods and compositions for long fragment read sequencing. The present invention encompasses methods and compositions for preparing long fragments of genomic DNA, for processing genomic DNA for long fragment read sequencing methods, as well as software and algorithms for processing and analyzing sequence data.

Methods of preparing a library of nucleic acid fragments tagged with oligonucleotide bar code sequences

Provided is a linker element and a method of using the linker element to construct a sequencing library, wherein the linker element consists of a linker A and a linker B, the linker A is obtained through the complementary pairing of a long nucleic acid strand and a short nucleic acid strand, the 5' end of the long strand has a phosphoric acid modification, and the 3&#x2032; end of the short strand has an enclosed modification, with enzyme sites in the short strand; and the linker B is a nucleic acid single strand, and the 3&#x2032; end thereof can be in a complementary pairing with the 5&#x2032; end of the long strand of the linker A. Using the linker element of the present invention for constructing a sequencing library ensures the linking directionality of the linkers while solving the problems of fragment interlinking, linker self-linking and low linking efficiency, and reducing the purification reaction between steps, shortening the linking time and reducing costs.

Linker element and method of using same to construct sequencing library

A method and reagent for constructing a nucleic acid double-linker single-strand cyclic library. The method comprises: breaking a nucleic acid into nucleic acid fragments; connecting a first linker sequence; producing by amplification a first product provided with the first linker sequence at either end, where a U nucleobase site is provided on primer sequences and a nicking enzyme recognition sequence is either provided or not provided on same, and a first affinity tag is provided on one of the primer sequences; using USER enzyme to cleave the first product; cyclizing the cleaved first product; treating the cyclization product with either a phosphatase or a nicking enzyme; using a solid-phase vector for combination with a cyclized molecule; performing a restrictive gap translation reaction; removing by digestion any portion that did not undergo the restrictive gap translation reaction; connecting a second linker sequence; producing by amplification a second product provided with the second linker sequence at either end; denaturing the second product, and cyclizing a single-strand nucleic acid molecule. The method allows an increase in the length of library insert fragments, a simplified library construction process, reduced library construction time, and reduced library construction costs.

Method and reagent for constructing nucleic acid double-linker single-strand cyclical library

This disclosure provides methods and compositions for tagging long fragments of a target nucleic acid for sequencing and analyzing the resulting sequence information in order to reduce errors and perform haplotype phasing, for example.

Multiple tagging of individual long DNA fragments

Reducing GC bias in DNA sequencing using nucleotide analogs

Methods and compositions for long fragment read sequencing

Growing community of inventors

Woodland, CA, United States of America

Andrei Alexeev