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Preparation method of modified DNA (deoxyribonucleic acid) hybridization probe for targeted hybrid capture

A hybrid capture and hybridization probe technology, applied in the field of genetic engineering, can solve the problems such as the lack of stability of the RNA hybrid capture probe, and achieve the effect of wide application range and lower production cost.

Active Publication Date: 2016-06-08
HANGZHOU LC BIOTECH
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Benefits of technology

This patented technology allows for less expensive ways to detect genetic material from different organisms or even within one cell type without affecting their functioning effectively. It also provides powerful tools with which it could help create new types of sequence data quickly by combining multiple fragments into larger pieces.

Problems solved by technology

This patented describes an improved method called Highly Efficient Panel System(HEPS), developed by NanoGen Inc., specifically designed to enable efficient and accurate discovery of new disease variants from their corresponding genomes. It uses advanced techniques such as massively parallel processing or bioinformatics to analyze multiple sequences simultanously without expensive equipment. Additionally, its goal was to discover novel drug targets through screening technologies like SELVERS™ or LCARIS®.

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  • Preparation method of modified DNA (deoxyribonucleic acid) hybridization probe for targeted hybrid capture
  • Preparation method of modified DNA (deoxyribonucleic acid) hybridization probe for targeted hybrid capture
  • Preparation method of modified DNA (deoxyribonucleic acid) hybridization probe for targeted hybrid capture

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Embodiment approach

[0021] A method for preparing a modified DNA hybridization probe for targeted hybridization capture, comprising the following steps:

[0022] (1) Obtain DNA templates containing target sequences and universal primer regions by preparing oligonucleotide pools in parallel.

[0023] (2) Use Taq polymerase and PCR primers containing dUTP to carry out PCR reaction, and introduce biotin-labeled ATP and methylated dm during PCR 5 CTP; the PCR primer is a pair of primers, including a forward primer and a reverse primer, the sequence of the forward primer is 5'-GAGCTTCGGTTCACGCAATG-3' (SEQ ID No.1), and the sequence of the reverse primer is 5'-UGCCUAGGACCGGAUCAAC-3 '(SEQIDNo.2), forward primer and reverse primer were synthesized by Shanghai Sangong. In the PCR reaction system, the final concentration of dUTP-containing PCR primers was 0.5 μM.

[0024] (3) The PCR product was purified by magnetic bead enrichment method.

[0025] (4) The purified PCR product was cleaved with USER and ...

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Abstract

The invention discloses a preparation method of a modified DNA (deoxyribonucleic acid) hybridization probe for targeted hybrid capture. The preparation method mainly includes preparing a DNA template prior to PCR (polymerase chain reaction) amplification and purifying a PCR product; subjecting the purified PCR product to USER enzyme digestion and T4 DNA polymerase terminal blunting, wherein one of double-stranded DNA of an Lambda exonuclease hydrolysis product has 5'-phosphorylated single strand, so that bioti-labeled single-stranded DNA is obtained; purifying the bioti-labeled single-stranded DNA by a magnetic beads enrichment method to obtain the modified DNA hybridization probe. The preparation method of the modified DNA hybridization probe for targeted hybrid capture has the advantages that production cost is reduced greatly, and the preparation method is wide in application range including exon regions, intron regions, mitochondrion regions and the like of genomes of any species. The obtained DNA hybridization probe can be applied to construction and sequencing of next generation targeting libraries.

Description

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Claims

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Application Information

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Owner HANGZHOU LC BIOTECH
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