High-throughput multi-LDR parting kit for detecting farmland ecological environment

An eco-environmental, high-throughput technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve problems such as difficult to achieve high-throughput detection

Inactive Publication Date: 2010-12-22
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a high-throughput multiple LDR typing kit for detecting the ecological environment of farmland, so as to overcome the current common typing kits that can only reveal the predation rules of a small number of prey in the field, and it is difficult to achieve high-throughput detection problem

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  • High-throughput multi-LDR parting kit for detecting farmland ecological environment
  • High-throughput multi-LDR parting kit for detecting farmland ecological environment
  • High-throughput multi-LDR parting kit for detecting farmland ecological environment

Examples

Experimental program
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Effect test

Embodiment 1

[0109] (1) Amplification of the DNA of the tarantula and the DNA of the prey with universal primers

[0110] (1) Mix the standard plasmids of 16 species at equal concentrations, and the total concentration is 10 pg / μL;

[0111] (2) Universal primers

[0112] Upstream: 5′-GGTCAACAAATCATAAAGATATTGG-3′;

[0113] Downstream: 5′-TAAACTTCAGGGTGACCAAAAAAATCA-3′;

[0114] (3) Amplification system (10μl):

[0115] Upstream primer 0.5μM

[0116] Downstream primer 0.5μM

[0117] 10×Buffer 1μl

[0118] dNTP 0.2mM

[0119] Mg 2+ 2.0mM

[0120] Taq polymerase 0.1μl (0.5Unit)

[0121] Add water to 10μl;

[0122] (4) Amplification program:

[0123] 95℃15min

[0124] 94℃30s→50℃1min→72℃2min(35cycles)

[0125] 72℃7min→4℃forever

[0126] (2) Ligase detection reaction (LDR) between the DNA of the tarantula and the DNA of the prey

[0127] (1) Use the PCR product amplified by the universal primer as a template for LDR signal detection

[0128] (2) LDR system (10μl):

...

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Abstract

The invention relates to a high-throughput multi-LDR parting kit for detecting farmland ecological environment, comprising mitochondria COI gene universal primers of 16 kinds of species in the farmland, specific probe sequences of the mitochondria COI gene universal primers of the 16 kinds of species in the farmland, and a reagent for PCR (polymerase chain reaction) proliferation and LDR (light dependent resistor) detection programs, wherein the mitochondria COI gene universal primers have the upstream of 5'-GGTCAACAAATCATAAAGATATTGG-3' and the downstream of 5'-taaacttcagggtgaccaaaaaatca-3'. In the invention, the kit effectively proliferates all target species sequences for the COI gene universal primers, and ensures that each target sequence and a low-copy template can be detected; the specific probe of each target species further ensures the specificity so as to totally distinguish each target sequence to realize multi-parting fixed quantity, thus the change of a farmland predation structure is known and the farmland ecological environment is maintained.

Description

technical field [0001] The invention belongs to the field of multiple LDR typing kits, in particular to a high-throughput multiple LDR typing kit for detecting farmland ecological environment. Background technique [0002] Multiple species in the field are extremely important to the farmland ecosystem, and it can provide guidance for which trophic relationship is more important. The detection and classification of multiple species in the field is very important for the study of ecosystem functions. At present, a number of detection and typing methods for multiple species in the field have been established at home and abroad, but these methods generally have weaknesses such as low throughput, difficult operation, cumbersome, and less information, which hinders research institutions from analyzing multiple species in the field. Carry out effective research and testing. The existing methods for detecting multiple species in the field are mainly divided into two types, one is t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 赵丽亚李凯田俊策叶恭银陈强周宇荀肖君华
Owner DONGHUA UNIV
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