Composite Primer Set for Cattle STR Identification

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Solution Overview

Problem

Current methods for cattle species identification and kinship analysis are hindered by high genealogical error rates, poor genotyping of STR loci, and the prevalence of dinucleotide repeats leading to low cumulative non-paternity exclusion rates, necessitating the development of a more effective kit for forensic identification.

Innovation Solution

A composite primer set and kit for identifying polymorphic markers in cattle, utilizing 13 STR loci including TGLA126, BT66, BT165, ETH10, and others, with optimized primer concentrations and fluorescent labeling for multiplex PCR amplification, enabling simultaneous amplification and genotyping of these markers.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If dinucleotide repeat STR loci are used for cattle identification, then amplification is achieved, but high stutter peaks are generated reducing identification accuracy

Engineering Contradiction:

Improveidentification accuracyVSAvoidstutter peaks

Core Design Contradiction:

ReliabilityVSObject-generated harmful factors

Solution Approach 1:

The patent extracts and eliminates the harmful dinucleotide repeat sequences that cause stutter peaks by selecting only tetranucleotide repeat STR loci for the identification system, thereby removing the source of identification errors while maintaining amplification capability

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent changes the fundamental parameter of STR locus selection from dinucleotide repeats to tetranucleotide repeats, which fundamentally alters the amplification characteristics and eliminates stutter peak generation while improving identification reliability

Inventive Principle:

Principle #35Parameter changes

2Device complexity

If fewer STR loci are used, then the identification system is simpler, but the cumulative non-paternity exclusion rate decreases

Engineering Contradiction:

Improvenumber of STR lociVSAvoidcumulative non-paternity exclusion rate

Core Design Contradiction:

Device complexityVSMeasurement precision

Solution Approach 1:

The patent combines 13 carefully selected tetranucleotide repeat STR loci into a single comprehensive identification system, achieving both high cumulative non-paternity exclusion rate (0.99999999997) and practical operability through optimized multiplex PCR protocols

Inventive Principle:

Principle #5Merging (Combining)

Solution Approach 2:

The patent creates a composite STR locus system comprising 13 specific tetranucleotide repeat loci (TGLA126, BT66, BT165, ETH10, CSM0113, INRA005, BT54, BT61, INRA023, BM2113, G18833, UMN0929, INRA063) that work synergistically to achieve extremely high identification accuracy

Inventive Principle:

Principle #40Composite materials

3Reliability

If commercialized STR kits are used for cattle identification, then species identification capability is provided, but genotyping effect is poor and cost is high

Engineering Contradiction:

Improvespecies identification capabilityVSAvoidgenotyping effect

Core Design Contradiction:

ReliabilityVSMeasurement precision

Solution Approach 1:

The patent develops a cost-effective alternative to expensive commercialized STR kits by using readily available reagents and standard PCR equipment, achieving superior genotyping effect while significantly reducing costs for forensic identification applications

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The solution provides a highly sensitive and specific forensic detection kit capable of accurate species, individual, and kinship analysis, reducing genealogical errors and increasing the cumulative non-paternity exclusion rate, suitable for judicial expertise and criminal investigations.

Implementation Method 1

a composite primer set for identifying polymorphic markers of cattle is provided, including: a nucleotide sequence of an upstream primer for amplifying TGLA126 gene shown in SEQ ID NO: 1, and a nucleotide sequence of a downstream primer for amplifying the TGLA126 gene shown in SEQ ID NO: 2

Methodology Applied

Scientific EffectPolymerase chain reaction (PCR):

Implementation Method 2

utilizing 13 STR loci including TGLA126, BT66, BT165, ETH10, and others, with optimized primer concentrations and fluorescent labeling for multiplex PCR amplification

Methodology Applied

Scientific EffectFluorescence: Fluorescence

Data Source

PatentUS20240360520A1Composite primer set and kit for identifying polymorphic markers of cattle and applications thereof

Publication Date: 2024.10.31 FUDAN UNIVERSITY

AI summary

A primer composition, a kit and a method for detecting polymorphic STR markers based on capillary electrophoresis and applications thereof are provided, which are used to simultaneously amplify 13 STR loci on cattle genome. The primer composition includes one or more pairs of primers with sequences as shown in SEQ ID NO: 1˜26. Developmental validation indicated that the kit is of high sensitivity, fine specificity, strong stability and high accuracy. This provides an effective tool for species identification, individual identification and parental analysis of cattle samples in civil disputes and illegal crimes.