Mgi Tech Co., Ltd.

Bgi-shenzhen Co., Limited

Bgi Shenzhen

Chunyu Geng

Hui Fu Jiang

Wenwei Zhang

Yuan Jiang

Radoje Drmanac

Xia Zhao

Lingyu He

Jing Guo

Kai Tian

Huaiqian Xu

Rongrong Guo

Xiaojun Ji

Shujin Fu

Yaqiao Li

Hongyan Han

Ruoying Chen

Handong Li

Jing Li

Dennis G Ballinger

Andrei Alexeev

He Zhang

Yingxin Zhang

Yanyan Zhang

Jing Yu

Zhicheng Li

Kailong Ma

Yanning Zhu

Wenhao Wu

Hui Jiang

Guangying Guo

Xiaoshan Su

Shaohua Li

Fanzi Wu

Y Tom Tang

Meiyan Li

Guanying Guo

Guangyi Fan

Specimen collector

Provided are a vesicular adaptor and a single-chain cyclic library constructed by using the adaptor. The library can be used for RNA sequencing and other sequencing platforms dependent on a single-stranded cyclic library, and has the advantage of high throughput sequencing, high accuracy and simple operations.

Vesicular adaptor and uses thereof in nucleic acid library construction and sequencing

Improved single molecule sequencing methods, compositions, and devices, are provided. In a first aspect, the present invention provides a multi-pass method of sequencing a target sequence using nanopore sequencing, the method comprising: i) providing a non-naturally occurring concatemer nucleic acid molecule comprising a plurality of copies of the target sequence; ii) nanopore sequencing at least three copies of the target sequence in the concatemer, thereby obtaining a multi-pass sequence dataset, wherein the multi-pass sequence dataset comprises target sequence datasets for the at least three copies of the target sequence; and iii) using the multi-pass sequence dataset to determine the target sequence.

Multi-pass sequencing

The present invention provides a primer for nucleic acid random fragmentation and a nucleic acid random fragmentation method. The primer consists of a plurality of upstream random primers and downstream random primers. The sequence composition of the upstream random primers is 5'-X-Y-3&#x2032;, and the sequence composition of the downstream random primers is 5&#x2032;-P-Y&#x2032;-X&#x2032;-close-3&#x2032;, wherein Y and Y&#x2032; are random sequences, X is all or part of sequences of a sequencing platform 5&#x2032; end adaptor, X&#x2032; is all or part of sequences of a sequencing platform 3&#x2032; end adaptor, P is phosphorylation modification, and close is close modification. The primer of the present invention adopts double random anchoring of both the upstream random primers and the downstream random primers, and a DNA sample can be randomly broken.

Primer for nucleic acid random fragmentation and nucleic acid random fragmentation method

Provided are a vesicular linker and a single-chain cyclic library constructed by using the linker. The library can be used for RNA sequencing and other sequencing platforms dependent on a single-stranded cyclic library, and has the advantages of high throughput sequencing, high accuracy and simple operations.

Vesicular linker and uses thereof in nucleic acid library construction and sequencing

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

Provided are a target region enrichment method based on multiplex PCR, and a reagent, the method comprising: connecting a first linker and a second linker respectively at two ends of a nucleic acid segment containing target regions to be enriched so as to obtain a linker-connected product; performing a PCR amplification on the linker-connected product using a first primer specifically bound to the first linker and a second primer specifically bound to the second linker to obtain an amplified product, the first primer or the second primer having a first affinity label; capturing a single strand having the first affinity label in the amplified product using a solid phase carrier; performing single primer linear amplification using a third primer with the captured single strand as a template; performing exponential amplification using the third primer and the first primer, with the linearly amplified product as the template, to obtain a product containing the target regions.

Target region enrichment method based on multiplex PCR, and reagent

Provided are a method for constructing a nucleic acid single-stranded cyclic library and the reagents used therein. By the combination of interruption via a transposase with a restricted nick translation reaction, the method realizes a simple and rapid nucleic acid single-stranded cyclic library construction.

Method for constructing nucleic acid single-stranded cyclic library and reagents thereof

Disclosed are a nucleic acid fragmentation method and a sequence combination. The method comprises the following steps: subjecting a denatured nucleic acid to annealing and an extension reaction by using a single-stranded 5'-end extension primer, wherein the single-stranded 5&#x2032;-end extension primer comprises a sequencing platform adaptor sequence of a 5&#x2032; end and a connected random sequence, and the random sequence is subjected to annealing on a random site of the denatured nucleic acid; and directionally connecting a double-stranded 3&#x2032;-end adaptor sequence to the 3&#x2032; end of the nucleic acid generated in the extension reaction, and carrying out denaturalization and purification to obtain a fragmented single-stranded nucleic acid with adaptor sequences on two ends.

Method and a sequence combination for producing nucleic acid fragments

Provided is a method of constructing a sequencing library using an adaptor element in a bubble shape. The bubble-shaped adaptor is ligated to a DNA fragment respectively at the 3'-terminal and the 5&#x2032;-terminal i.e., two bubble-shaped adaptors with same sequences are ligated in one step. The bubble-shaped adaptor-ligated product is then amplified with a primer complementary to the 3&#x2032;-terminal of the long-chain nucleic acid of the bubble-shaped adaptor, so as to replace the non-paired sequence in the short-chain nucleic acid.

Method of constructing sequencing library with bubble-shaped adaptor element

Provided in the present invention are a method for constructing a nucleic acid single-stranded cyclic library and reagent kit thereof. The method comprises the steps of using a transposase embedding complex to randomly break nucleic acids and connect a first linker; connecting a second linker at a gap; performing a first PCR reaction, wherein the 5' end of one of the primers has a first affinity tag, resulting in a product with two ends connected to different linker sequences; binding the product to a solid vector having a second affinity tag; degenerating and separating single strands having no affinity tag; and cyclizing the single strands.

Provided are an isolated oligonucleotide and a use thereof in nucleic acid sequencing, wherein the isolated oligonucleotide comprises a first strand, wherein the 5'-end nucleotide of the first strand has a phosphate group, and the 3&#x2032;-end nucleotide of the first strand is a dideoxynucleotide, and a second strand, wherein the 5&#x2032;-end nucleotide of the second strand does not have a phosphate group, and the 3&#x2032;-end nucleotide of the second strand is a dideoxynucleotide, wherein the first strand is longer than the second strand in length, and a double-stranded structure is formed between the first strand and the second strand.

Isolated oligonucleotide and use thereof in nucleic acid sequencing

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Shenzhen, China

Chunyu Geng