Primer-mediated cyclized constant-temperature nucleic acid rolling circle amplification method and kit

A rolling circle amplification and cyclization technology, applied in biochemical equipment and methods, microbial assay/inspection, etc., can solve the problem of not being able to be completed at one time
CN103255227AActive Publication Date: 2013-08-21QUICKING BIOTECH
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
QUICKING BIOTECH
Publication Date
2013-08-21

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Abstract

The invention discloses a primer-mediated cyclized constant-temperature nucleic acid rolling circle amplification method and a kit. The method comprises the following steps of: 1) designing an oligonucleotide sequence; 2) amplifying a single-stranded DNA (deoxyribonucleic acid) target template from target nucleic acid; 3) cyclizing the DNA target template through a DNA ligase; and 4) performing rolling circle amplification on the cyclized DNA target template. According to the method disclosed by the invention, a long padlock probe with high cost does not need to be synthesized, the length of the sequence of the target nucleic acid can be allowed to be longer, the rolling circle amplification can be performed on any sequence of the target nucleic acid, a solution can be provided for preventing a sample from being polluted by an amplified product, and a new era that gene detection enters applications in basic level units is opened.
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Description

technical field

[0001] The present invention relates to a nucleic acid sequence constant temperature amplification method and kit, more specifically, to a constant temperature nucleic acid rolling circle amplification method for primer-mediated circularization, that is, two primer-related sequences for preparing single-stranded DNA strands A method and a kit (Primer mediated Rolling Circle Amplification, PRCA) capable of initiating constant temperature rolling circle amplification with at least two primers connected to form a circle at the ends. Background technique

[0002] In 1985, the U.S. Cetus company used Polymerase Chain Reaction (Polymerase Chain Reaction, PCR) technology to realize the in vitro amplification of nucleic acid for the first time, which played a very important role in the development of modern molecular biology. A revolutionary initiative and milestone in the field of medicine. However, the PCR method still has deficiencies such as: the double-stranded...

Claims

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