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Longan detection kit and longan detection method

A technology for detection reagents and detection methods, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problem of easy confusion between longan and longli, achieve good application prospects and ensure food safety , the effect of easy operation

Inactive Publication Date: 2014-11-19
傅俊江
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problem that longan and longan are easily confused, the present invention provides four kinds of longan-specific nucleotide sequences and their uses, and also provides longan detection kits and detection methods

Method used

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  • Longan detection kit and longan detection method
  • Longan detection kit and longan detection method
  • Longan detection kit and longan detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 RAPD method specific amplification to obtain longan-specific SCAR markers

[0032] RAPD (Random Amplified Polymorphic DNA) - that is, random amplified polymorphic DNA, is mainly used for detection when the genomic information of animals and plants is unknown. RAPD technology relies on PCR, and selects 10 base single-stranded random primers to perform PCR amplification on the DNA of organisms, populations or species genomes to detect differences in genetic loci. Sequence-characterized Amplified Region (SCAR) markers (SCAR markers) were developed on the basis of RAPD technology. SCAR markers generally manifest as the presence or absence of amplified fragments, which is a dominant marker; but sometimes also manifest as length polymorphisms, which are co-dominant markers. As a SCAR marker for crop agronomic traits, it will play a huge role in molecular marker-assisted breeding and variety identification.

[0033] 1. Experimental method

[0034] (1) Extraction of ...

Embodiment 2

[0136] Example 2 Using the method of the present invention to specifically amplify the longan gene

[0137] 1. Experimental method

[0138] (1) According to the sequences shown in SEQ ID NO.1-4 obtained in Example 1, use PRIMER3 software to design primers for analysis, design 8 pairs of primers, and synthesize them.

[0139]

[0140] The specific primer sequences are as follows:

[0141] 2-1 Primer pair (SEQ ID NO.5-6):

[0142] LY2-1L: 5'-AACTGGAAGTCCCTGGTCCT-3'

[0143] LY2-1R:5'-ACAAGAGGCCCCAGTAAGGT-3'

[0144] 4-7 primer pairs (SEQ ID NO.7-8):

[0145] LY4-7L:5'-GGCGCCGGTATACTTTGTAA-3'

[0146] LY4-7R:5'-CTCGTAAGAGGATCCGTCCA-3'

[0147] 4-8 primer pairs (SEQ ID NO.9-10):

[0148] LY4-8L:5'-CCCCATCTGGTTGTAGCACT-3'

[0149] LY4-8R:5'-AGCCAGCTCAACCAACTCAT-3'

[0150] 5-2 Primer pair (SEQ ID NO.11-12):

[0151] LY5-2L:5'-TTTTAATGTTGGGCATTTGG-3'

[0152] LY5-2R:5'-GCTAACCGAGATCCACTAACG-3'

[0153] LY2-1-full primer pair (SEQ ID NO.13-14)

[0154] LY2-1L-full TAC...

Embodiment 3

[0187] Embodiment 3 Longan detection kit of the present invention

[0188] 1. Kit composition

[0189] This kit contains:

[0190] CTAB extraction buffer (200ml);

[0191] 2×Taq Master Mix, wherein the primers can be selected from SEQ ID NO.5~6, SEQ ID NO.7~8, SEQ ID NO.11~12, SEQ ID NO.13~14, SEQ ID NO.15~16 , a pair of primers or multiple pairs of primers (500 μl) among the primers shown in SEQ ID NO.17-18 and SEQ ID NO.19-20;

[0192] One tube (50 μl) of negative control template DNA (Longli DNA);

[0193] One tube (50 μl) of positive control template DNA (longan DNA);

[0194] Sterile ddH2O (1500 μl).

[0195] (1) CTAB extraction buffer

[0196] CTAB extraction buffer (100ml)

[0197] 2.0g CTAB (Hexadecyl trimethyl-ammonium bromide, hexadecyl trimethyl-ammonium bromide)

[0198] 10.0ml 1M Tris (pH8.0)

[0199] 4.0ml 0.5M EDTA (pH8.0)

[0200] 28.0ml 5M NaCl

[0201] 40.0ml H 2 o

[0202] 1g PVP 40 (polyvinyl pyrrolidone, polyvinylpyrrolidone, Mw40,000) was adjus...

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Abstract

The invention discloses four new nucleotide sequences of longan, and also discloses an application of a reagent for detecting the sequences in the preparation of a longanon detection reagent, as well as a longan detection kit and a longan detection method. The detection kit and the detection method provided by the invention can be used for accurately and effectively detecting the longan, is strong in specificity and short in consumed time, and has a good application prospect.

Description

technical field [0001] The invention relates to a detection kit and a detection method for longan. Background technique [0002] Longan (Dimocarpus longan Lour), commonly known as "longan", belongs to Sapindaceae Longan. It is a traditional medicinal and edible plant, mainly distributed in tropical and subtropical regions. As the birthplace of longan, China has a cultivation history of nearly a thousand years. In history, it is known as "longan" in the south and "ginseng" in the north. China is the country with the richest longan genetic resources and the most cultivated varieties in the world. [0003] Madman fruit, also known as longli, is poisonous, and its morphological characteristics are very similar to longan, which is easy to confuse, leading to poisoning of consumers, and there is a great potential safety hazard. It is urgent to provide a method to solve this problem. Contents of the invention [0004] In order to solve the problem of easy confusion between long...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 傅俊江杨璐全
Owner 傅俊江