Construction and application of general oligonucleotide sequence database
A technology of oligonucleotide and sequence library, which is applied in the direction of nucleotide library, library, chemical library, etc., can solve the problems that gene library group analysis cannot be performed, and genes with unknown sequences are not applicable, so as to reduce costs and improve experimental efficiency Effect
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Embodiment 1
[0018] Example 1: Construction of a general oligonucleotide probe library for multi-species transcriptomes such as figure 1 , figure 2 As shown, based on bioinformatics analysis and algorithm design, the present invention constructs a general oligonucleotide probe library for transcriptomes of multiple species, specifically involving 8bp and 9bp general probe libraries of 16 species. A class of oligonucleotide sequences that can be used as probes and primers is sorted according to their frequency of occurrence in the target sequence set, and further screened to obtain a statistically significant high-frequency candidate probe library. The distribution of each probe and different combinations of probes in the high-frequency probe library in the target sequence set is analyzed by a greedy algorithm. Achieve the maximum coverage of species transcript sequences with the least number of universal probes, and ensure the uniformity and distribution density of probes in each transcr...
Embodiment 2
[0021] Example 2: Construction of a general oligonucleotide sequence library for multi-species genomes figure 1 , figure 2 With the flow and method shown, the present invention constructs a general oligonucleotide sequence library for the genomes of multiple species. It specifically involves the 8bp and 9bp general sequence libraries of 16 species that have been sequenced. For each species, a certain proportion (can be 5%-50%) of the whole genome sequence is randomly selected as the target sequence set, according to figure 1 The process of screening the candidate high-frequency probe library for the target sequence set. The greedy algorithm for analyzing the genome sequence is designed as follows: for each candidate probe or probe combination, the matching of multiple or different probes that appear in a continuous region (generally 15-50 bases) of the target sequence, are calculated only once. pass figure 2 Schematic flow of the greedy algorithm to analyze and obtain a...
Embodiment 3
[0024] Embodiment 3: Utilize the general oligonucleotide sequence library to carry out the combinatorial design of capture probe Liquid chips based on magnetic beads (magnetic microspheres) or other substrates. Such as image 3 In the experimental process, for the target gene (or a group of genes) that needs to be separated and purified, the optimal sequence arrangement probe set can be designed in the general probe library p 1 ,p 2 ,...,p i . After the nucleic acid sequence library sample is combined with the capture probe and eluted, it can be split into two secondary libraries with the probe binding site and without the probe binding site; the initial library sequence is passed through all Screening at all levels of capture probes can finally separate and purify the target gene (or a group of genes). Taking the 8bp capture probe design strategy as an example, design a set of 3 universal capture probes, the total length of which is equivalent to designing a 24bp probe; ...
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