DNA tags, PCR primer and application thereof

A technology for labeling and amplification products, applied to DNA labels, PCR primers and their application fields, can solve the problems of limited resolution, difficulty in number, and inability to further detect minor differences in the primary structure of nucleic acids, so as to improve resolution and sequencing. High throughput, accurate and reliable STR detection results

Active Publication Date: 2016-02-10
TIANJIN MEDICAL LAB BGI +1
View PDF3 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to technical limitations, this method also has certain defects, mainly including: (1) Due to the mutual interference of fluorescent markers and the limitations of capillary length and imaging technology, the number of analyzed STR loci has been limited. It is difficult to further greatly improve; (2) Since the object of analysis is the length of each fragment, it is impossible to further detect the slight difference in the primary structure of the nucleic acid that makes up the fragment, thus limiting the resolution of detection; (3) There are invalid alleles , resulting in differences in the determination results of some loci that may occur in different kits; (4) the interference of Stutter peaks (small peaks that sometimes appear before the main peak in fragment analysis), especially when there are mixed samples; (5) Sanger method For STR typing, due to throughput, cost and other reasons, it is not conducive to the construction of STR typing database
[0004] Therefore, the current detection and typing methods of STR loci still need to be improved

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • DNA tags, PCR primer and application thereof
  • DNA tags, PCR primer and application thereof
  • DNA tags, PCR primer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] Adopt the method of the present invention, carry out STR gene loci typing to 190 samples, concrete steps are as follows:

[0091] 1. Sample extraction

[0092] DNA was extracted from 190 dried blood slices with 5% chelex (chelex-100 brand BIO-RAD). After the extraction, the dried blood slice extraction product with a diameter of 3 mm was obtained, which was used as a template in the next step of PCR amplification.

[0093] 2. PCR amplification

[0094] The 190 copies of DNA obtained in step 1 were numbered 1-190 in turn, and divided into 2 groups evenly (STR-1 group: numbering 1-95; STR-2 group: numbering 96-190). According to the sequence (SEQIDNO:96-135) of each primer of the primer set (comprising 20 forward primers and 20 reverse primers) that is used to amplify STR gene, design a set of labels, altogether 95 (SEQIDNO:1 -95). Add each designed tag to the 5' end of the sequence of each primer in the primer set to obtain 95 tag primer sets, where each tag primer s...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses DNA tags, a PCR primer and application thereof. The DNA tags of one group are chosen from nucleotides shown as in SEQ ID NO:1-95 and can be used for building a nucleic acid sequencing library to accurately differentiate the nucleic acid sequencing library. The DNA tags and the PCR primer are utilized to build a tagged PCR primer, and by utilizing the tagged PCR primer, STR detection of at most 95 DNA samples can be realized at one step according to a method for determining preset STR gene locus genotype of various DNA samples.

Description

technical field [0001] The present invention relates to the technical field of nucleic acid sequencing and typing, in particular, to DNA tags, PCR primers and applications thereof, more specifically, to a set of DNA tags, a set of PCR primers, a set of tagged PCR primers, and construction of nucleic acid sequencing libraries The method, the method for determining the genotype of the predetermined STR locus of a DNA sample, the kit for constructing a nucleic acid sequencing library and the system for determining the genotype of the predetermined STR locus of a DNA sample. Background technique [0002] STR locus sequence, also known as short tandem repeat sequence (Shorttandem repeat) is a kind of widely distributed genetic markers in the human genome, usually composed of 2-7 core bases, and the number of repeat units is different, resulting in different individuals in the same locus. alleles. The allele types can be typed by techniques such as silver staining, fluorescent la...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C40B50/06C12Q1/68C12M1/34
Inventor 张俊青刘玉强程秀穆豪放陈祖煜吴仁花易鑫杨玲
Owner TIANJIN MEDICAL LAB BGI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap