Construction method of dated bloodstain miRNA high-throughput sequencing library

A sequencing library and construction method technology, applied in the field of molecular biology, can solve the problems of high amount of miRNA templates, inability to apply trace blood stains, no reports on miRNA library construction methods and sequencing methods, etc., and achieve the effect of increasing lysis time
CN108977435AActive Publication Date: 2018-12-11BEIJING CENT FOR PHYSICAL & CHEM ANALYSIS
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
BEIJING CENT FOR PHYSICAL & CHEM ANALYSIS
Publication Date
2018-12-11

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Abstract

The invention discloses a construction method of a dated bloodstain miRNA high-throughput sequencing library. The method comprises the steps of pretreatment of a dated bloodstain sample, extraction ofmiRNA and construction of a miRNA sequencing library, wherein the library construction includes: (1) connection of miRNA 5' and 3' junctions; (2) reverse transcription; (3) PCR (Polymerase Chain Reaction) amplification; (4) agarose gel electrophoresis on a PCR amplification product, gel cutting and recovery of fragments of 145 to 160 bp to obtain the sequencing library. By adopting the method disclosed by the invention, construction of the dated bloodstain miRNA library can be finished within 48 hours, and only a 5cm<2> dated bloodstain (containing about 10 to 100ng RNA) sample is needed at least. Reuse of dated forensic evidence is facilitated, the reuse of dated forensic evidence appears in the field of forensic medicine for the first time, and strong technical support is provided for expanding the application of miRNA in forensic medicine.
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Description

technical field

[0001] The invention relates to the field of molecular biology, in particular to a method for constructing an old bloodstain miRNA high-throughput sequencing library. Background technique

[0002] Individual identification is an important task of forensic evidence, and it is used to reveal individual identity, investigate and solve cases, and an important link in compensation. Traditional individual identification methods are based on genomic DNA short tandem repeats (Short Tandem Repeats, STRs), single nucleotide polymorphisms (Single Nucleotide Polymorphisms, SNPs) analysis and so on. These methods have high requirements on the amount and integrity of templates, and when the amount of templates is too small or degraded, the accuracy of the detection results will be affected. At the same time, it is also impossible to judge the tissue, body fluid source and individual characteristics of the sample.

[0003] Early individual identification used the method o...

Claims

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