Sequencing library, and preparation and use thereof
AssigneeAgricultural Genomics Institute at Shenzhen China Academy of Agricultural Sciences
InventorsJue Ruan, Kaile Wang
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Abstract
The present invention discloses a sequencing library comprising a nucleotide sequence. The sequence comprises a linker sequence and two target sequences. Two ends of the linker sequence are respectively linked to the target sequences and the two target sequences are direct repeat sequences. The present invention further discloses preparation and use of the sequencing library. The present invention overcomes the high error rate problem of current DNA sequencing technologies, especially in a way of very low coverage bias, and can be used to detect low frequency mutations in different kinds of samples.
Claims8
A method for preparing a polynucleotide for sequencing, the polynucleotide comprising a linker and exactly two target sequences, wherein the two target sequences are respectively linked to two ends of the linker, the nucleotide sequence of each of the target sequences is different from the nucleotide sequence of the linker, and the two target sequences are direct repeat sequences, the method comprising:
- obtaining a double-stranded circular polynucleotide having nicks or gaps in both strands;
- and
- subjecting the double-strands circular polynucleotide with nicks or gaps in both strands to strand displacement amplification, forming the polynucleotide for sequencing.
A method for preparing a polynucleotide for sequencing, the polynucleotide consisting of a linker and exactly two target sequences, wherein the two target sequences are respectively linked to the two ends of the linker, the nucleotide sequence of each of the target sequence is different from the nucleotide sequence of the linker, and the two sequences are direct repeat sequences, the method comprising:
- obtaining a double-stranded circular polynucleotide having nicks or gaps in both strands;
- and
- subjecting the double-strands circular polynucleotide with nicks or gaps in both strands to strand displacement amplification, forming the polynucleotide for sequencing.
A method for preparing a sequencing library, comprising the steps of:
- obtaining a double-stranded circular polynucleotide having nicks or gaps in both strands;
- and
- subjecting the double-strands circular polynucleotide with nicks or gaps in both strands to strand displacement amplification, forming polynucleotide, subjecting the polynucleotide to a standard high-throughput sequencing library preparation;
- wherein the polynucleotide comprises a linker and two target sequences, two ends of the linker are respectively linked to the target sequences, and the two target sequences are direct repeat sequences.
Dependencies are inferred from the claim text.