Isothermal oligonucleotide library amplification method applied to DNA data storage

Abstract

The invention relates to the field of biotechnology, and develops a method for homogenizing concentrations of different oligonucleotides in an oligonucleotide library and amplifying the oligonucleotide library at a normal temperature. The method can be applied to DNA data storage. The method can homogenize the concentrations of the different oligonucleotides, and can also improve the purity of oligonucleotide fragments. According to a strand displacement amplification technique, the oligonucleotide library can be amplified at a constant temperature, and an amplified product is ssDNA with a phosphate group, so that great convenience is brought to library preparation in subsequent second-generation sequencing. Nickase required in an SDA reaction has a long recognition sequence, which makes data coding easier. In addition, the amplification method is a linear amplification method in which a most original template is used for amplification, so that mutation is not amplified as compared toPCR, and multiple rounds of amplification can be performed.

Description

technical field [0001] The invention relates to the field of biotechnology, and more specifically relates to an oligonucleotide library constant temperature amplification method applied to DNA data storage. Background technique [0002] Oligo pool is an important reactant in genomics, biophysics, bionic biology and biotechnology applications; oligo pool can be used as primers and probes in the amplification, enrichment, detection and sequencing of biological nucleic acids. Researchers use Oligo pool to achieve gene assembly and synthesis; use it as a probe for SNP genotyping; as scientists discover that DNA can be used as a storage medium to store information on a large scale, researchers use chip synthesis Oligo pool combined with DNA data storage Store various information in DNA. [0003] At present, people are generating data at an exponential growth rate, and its growth rate has exceeded the growth rate of storage hard disks, making human beings have higher demands on s...

AI Technical Summary

Problems solved by technology

[0006] However, this technology has the following disadvantages: 1) Precise temperature and cycle control are required, and heat is bound to be released during this process, which limits the application of DNA data storage in the future (because heat is also a factor that affects CPU performance) important factor); 2) The PCR instrument, which can accurately control the temperature and cycle, is required, which relatively increases the cost; 3) The temperature of the PCR process is relatively high, and the high temperature will speed up the decay rate of DNA and reduce the DNA data. Storage time; 4) The PCR process has a certain sequence preference, which will cause some data loss; 5) The PCR product is a double-stranded product, and its 5' end has no phosphate group

Although the stoichiometric oligonucleotide library purification method (SNOP) introduced by Zhang, DY, etc. can achieve simultaneous purification, the preliminary design and chemical modification are more complicated.

Images