Abstract: The present invention discloses a probe applicable to high-throughput sequencing and a method of enriching a target region using the probe. The probe includes, in an order from a 5? end, a ligation arm, a masking sequence 1, a UID sequence 1, an Illumina Tag1 sequence, a dU region, an Illumina Tag2 sequence, a UID sequence 2, a masking sequence 2 and an extension arm; wherein the masking sequence 1, the masking sequence 2, the dU region, the UID sequence 1, and the UID sequence 2 may be 0 bp. When sequencing is performed by using a library built by the probe of the present invention, it is not necessary to add a sequencing primer additionally, and adopting the method of the present invention may directly use an RNA as a template to capture a target region, so as to simplify the experiment procedure.

Abstract: A multi-position double-tag adapter set for detecting gene mutation and preparation method therefor and application thereof, the multi-position double-tag adapter set comprising a double-tag adapter A, a double-tag adapter B and a double-tag adapter C. The double-tag adapter A, the double-tag adapter B and the double-tag adapter C are obtained respectively by hybridizing an adapter primer P5 with an adapter primer P7-A, an adapter primer P7-B and an adapter primer P7-C 5? ends of which are all modified with biotin. Using the multi-position double-tag adapter set, the mutation rate of 1×10?5 genes may be accurately detected and the sensitivity of gene mutation detection may be effectively improved. A plurality of mutation sites of a plurality of genes may be detected by one-time sequencing in combination with throughput of high-throughput sequencing.

Abstract: A multi-position double-tag connector set for detecting gene mutation and preparation method therefor and application thereof, the multi-position double-tag connector set comprising a double-tag connector A, a double-tag connector B and a double-tag connector C. The double-tag connector A, the double-tag connector B and the double-tag connector C are obtained respectively by means of synthesizing a connector primer P5 with a connector primer P7-A, a connector primer P7-B and a connector primer P7-C of 5? ends which are all modified with biotin. Using the multi-position double-tag connector set, the mutation rate of 1×10?5 genes may be accurately detected and the sensitivity of gene mutation detection may be effectively improved. A plurality of mutation sites of a plurality of genes may be detected by one-time sequencing in combination with throughput of high-throughput sequencing.

Abstract: Loop-shaped primer used in nucleic acid amplification is an oligonucleotide with 3-20 bases in both 3? and 5? ends which can be combined together to form a double-strand under appropriate conditions, resulting in the primer forming a stem-loop structure. The double-stranded structure is opened and the stem-loop structure dissolves when the primer recognizes and hybridizes with the target sequence. If the target sequence is not present the primer can form a stem-loop structure automatically by self-annealing. The primer can comprise a universal tag sequence or not. Together with universal tag sequence primer the primer comprising the universal tag sequence can be used for a second round of amplification. The primer has high specificity and does not form a primer dimmer. The primer is easy to design and is suitable for measuring gene expression and detecting features of nucleic acids such as SNPs and rare mutations.

Abstract: The present invention is intended to provide a method for predicting risk of hepatocellular carcinoma (HCC) in hepatitis B virus (HBV)-infected patients with a high accuracy. More specifically, the invention provides a method for detecting eight mutations of HBV genome associated with predisposition to HCC, comprising: C1653T, A1762T, G1764A, T1674C, T1753C, C3116T, T53C and A1846T mutations, and primers and probes sets used thereof consist of SEQ ID NO: 1-SEQ ID NO: 24.