Novel gene cloning method independent of enzymes

A cloning method and a new technology, applied in the field of genetic engineering, can solve the problems of complete degradation or degradation of less than 15 nucleotides, the inability to effectively control the length of single-strand overhangs, and the difficulty of cloning target genes, etc., to achieve simple cloning Quantity, elimination of false positive clones, and improvement of lysis efficiency

Inactive Publication Date: 2019-11-26
苏州博睐恒生物科技有限公司
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Benefits of technology

This patented innovation describes an improved way to modify or remove certain parts (called “thiol groups”) from biological molecules called nucleic acids. These modifications are made with special techniques like chemistry, radiation, etc., making them easier than traditional methods such as enzyme cutting. They allow precise manipulation of these modified areas without affecting their original properties. By breaking down this process into individual strands they create more efficient ways to study how different types of cells work together. Overall, this new technique allows researchers to efficiently manipulate complex organisms while maintaining good quality results.

Problems solved by technology

Technological Problem addressed in this patents relates to improving the accuracy and reliability of gene clones by introducing specialized sequences into their structure during molecular biology procedures like PCR amplification. Existing techniques require complicated steps that can lead to loss of desired parts due to errors made during manipulation. Therefore, new ways to improve these processes would provide technical benefits with improved effectiveness and precision.

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  • Novel gene cloning method independent of enzymes
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Embodiment Construction

[0018] The technical solutions in the embodiments of the present invention will be described clearly and completely below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative work shall fall within the protection scope of the present invention.

[0019] See figure 1 , Provide a novel gene cloning method that does not rely on enzymes, including the steps:

[0020] (1) Design and synthesize the primer sequence (oligonucleotide fragment) used to amplify the target gene and the plasmid vector. The characteristics of the primer sequence are: (a) There is an intermittent thiomodification between the 1-25 position at the 5'end The phosphodiester bond is 5-8 bases apart between adjacent thiomodifications; (b) The base sequence between the 5'end of the forward primer of the amplifi

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Abstract

The invention discloses a novel gene cloning method independent of enzymes. The novel gene cloning method comprises the following steps of amplifying a target gene and a plasmid vector with a primer;performing heat treatment on target gene amplification products and plasmid vector amplification products with iodine; performing hybridization pairing on target gene DNA fragments and plasmid vectorDNA fragments after heat treatment, to form a recombinant plasmid with a notch, containing the target gene; and transforming escherichia coli with the recombinant plasmid with a notch, containing thetarget gene, and repairing the notch of the recombinant plasmid to form a complete recombinant plasmid containing the target gene. In the manner, the best sulfo modifying and single-strand end lengthcontrol strategy is adopted, the positive cloning rate of the target gene is high, the operation flux is high, and the method is suitable for simultaneous cloning of a plurality of target genes.

Description

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Claims

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Application Information

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Owner 苏州博睐恒生物科技有限公司
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