The invention discloses a method and a kit for enriching a DNA (deoxyribonucleic acid) target area by anchoring nested multiplex PCR (polymerase chain reaction). The method comprises the following steps: performing end repairing and linker sequence connection on DNAs, wherein linker sequences comprise a long sequence and a short sequence, which are mutually complementary, the long sequence is complementary to the short sequence at a position close to a connecting end, and the other part comprises a segment of tag sequence; performing a first round of multiplex PCR on a connection product, wherein an upstream primer is an anchoring product of which at least a part is the same as the tag sequence, and a downstream primer comprises multiple sequences specifically bound to the target area; performing a second round of multiplex PCR on a product obtained by the first round of multiplex PCR, so as to finally obtain a target area library, wherein an upstream primer is an anchoring primer, and a downstream primer comprises multiple sequences of which 5'-ends have sequencing primer sequences and 3'-ends are specifically bound to the target area. The method has the characteristics of high speed, low cost and high throughput.

Method and kit for enriching DNA (deoxyribonucleic acid) target area by anchoring nested multiplex PCR (polymerase chain reaction)

The invention provides method and device for determining a sample source of reading segments in mixed sequencing data. The mixed sequencing data consist of a plurality of reading segments. The method comprises the following steps: respectively marking a plurality of nucleic acid samples by using a plurality of labels so that each nucleic acid sample has at least one or more labels so as to differentiate from other nucleic acid samples, and recording the corresponding relationship of each nucleic acid sample and the labels of the nucleic acid sample; mixing the plurality of nucleic acid samples and sequencing the mixed nucleic acid samples to obtain the mixed sequencing data; comparing the mixed sequencing data with a reference sequence to obtain a comparison result and screening the reading segments which are incompletely matched with the reference sequence from the comparison result, wherein the length of the unmatched part is not less than the length which is 1bp less than the label; and determining the nucleic acid samples in which the reading segments are originated in the mixed sequencing data according to the corresponding relationship between the information and labels of the screened reading segments and the samples.

Method and device for determining sample source of reading segments in mixed sequencing data

The invention discloses an adapter for next-generation sequencing. The adapter disclosed in the invention is composed of a single-stranded DNA A and a single-stranded DNA B or composed of a single-stranded DNA C and a single-stranded DNA D. The single-stranded DNA A, from terminal 5' to terminal 3', is as shown in a formula (I), i.e., A1-A3-A2, wherein each nucleotide of A3 is A, T, C or G; the single-stranded DNA B, from terminal 3' to terminal 5', is as shown in a formula (II), i.e., B1-B2, wherein A2 and B2 are complementary, A1 and B1 are not complementary, A3, B1 and A1 are not complementary, the sequences of A1 and A2 are different, and the sequences of B1 and B2 are different; the single-stranded DNA C is composed of A1 and A2; and the single-stranded DNA D is composed of B1, B2 and A3. Experimental results show that the adapter provided by the invention can easily and efficiently reduce false positive mutation in next-generation sequencing so as to realize more sensitive detection of low-frequency mutation in heterogeneous mixed samples like heterogeneous samples and chimera samples of tumors.

Adapter for next-generation sequencing

A sequencing data result analysis method and apparatus, and a sequencing library construction and sequencing method The sequencing data result analysis method comprises: acquiring a sequencing data result of a sequencing library (S101), with the sequencing library comprising a plurality of mixed samples, each of the samples corresponding to a label sequence combination, and different samples corresponding to different label sequence combinations, wherein each label sequence combination comprises a plurality of label sequences, the sequencing data result is a sequencing fragment set obtained by sequencing the plurality of mixed samples, and the sequencing fragment set comprises a plurality of disordered sequencing fragments; determining a label sequence combination of each of the sequencing fragments (S102); and determining, according to the label sequence combination of each of the sequencing fragments, a sample corresponding to each of the sequencing fragments (S103) The method solves the technical problems of a relatively low efficiency and relatively high costs caused by the artificial discrimination against samples needing to be performed by scientific and technological workers with a technical background for a sequencing data result in the relevant art

Sequencing data result analysis method and apparatus, and sequencing library construction and sequencing method

The invention provides a blood plasma DNA library building method and kit. The library building method comprises the following steps: repairing the tail end of blood plasma DNA and adding A to obtainrepaired DNA; and sequentially carrying out joint connection and PCR amplification on the repaired DNA by adopting a joint with a unique tag sequence to obtain a sequencing library of the blood plasmaDNA. The sequencing library of the blood plasma DNA is constructed by joint connection of the blood plasma DNA with unique tag sequence after repairing and A addition. By the formed library, repeatsgenerated by library-building amplification or sequencing amplification according to the unique tag sequence are conveniently distinguished from real repeat sequences in the blood plasma DNA when sequencing data are analyzed at a later stage, then amplification repeats are removed, repeats exist in blood plasma fragments DNA retains, thus, the effective data size of blood plasma-derived DNA is increased, and the blood plasma DNA library building efficiency is improved to a certain extent.

Blood plasma DNA library building method and kit

Multi-sample mixed sequencing method and kit

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Multi-sample mixed sequencing method and kit