Method of using genes for species identification of plants

A gene pairing and plant technology, applied in the field of genetic engineering, can solve problems such as easy confusion, achieve the effect of improving accuracy and shortening identification time

Inactive Publication Date: 2016-07-06
SHANGHAI PASSION BIOTECHNOLOGY CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

If identified by morphological methods, it is ext...

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  • Method of using genes for species identification of plants
  • Method of using genes for species identification of plants
  • Method of using genes for species identification of plants

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Embodiment

[0027] This embodiment relates to a method for identifying the species of plants using genes; specifically, it includes the following steps:

[0028] 1. Genome Extraction

[0029] 1. Weigh 200 mg of fresh plant tissue (if freeze-dried tissue is used, the amount of tissue should be halved). Cut into small pieces and put them in a mortar; add liquid nitrogen to freeze the tissue completely, then quickly and vigorously grind to powder. Liquid nitrogen should be added intermittently during grinding to prevent the tissue from melting. After the grinding is complete, put the mortar in a 56°C water bath until the sample powder just begins to melt;

[0030] 2. Add 350 μl PBS and 0.9 μl RNaseA stock solution, and grind vigorously for 30 seconds;

[0031] 3. Transfer 350 μl of the ground homogenate to a 2ml centrifuge tube, if the volume of the homogenate is less than 350 μl, add PBS to 350 μl;

[0032] 4. Add 150 μl buffer C-L and 20 μl proteinase K; immediately vortex for 1 min to ...

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Abstract

The invention discloses a method for identifying the species of plants by using genes; it comprises the following steps: extracting the genome DNA of the plant sample to be tested; using the genome DNA of the plant sample to be tested as a template, using primers designed for the rbcL gene sequence and matK The primers designed for the gene sequence were amplified by PCR; the PCR amplified products were recovered to obtain the target DNA; the target DNA was sequenced using the Sanger dideoxy chain termination method; the sequencing results were compared with the sequences in the public database, according to Homology identifies the test plant to a genus or species. The present invention identifies the species of the plant from the molecular level, which is more accurate than the morphological identification; there is no too much requirement on the integrity of the plant form, as long as there is a little tissue of the plant itself, DNA can be extracted and identified by the present invention.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a method for identifying the species of plants by using genes. Background technique [0002] DNA barcoding technology is a new biological identification system created by using standard, sufficiently variable, easy-to-amplify, and relatively short DNA fragments within species for specificity and inter-species diversity, so as to realize the Rapid automatic identification of species. The use of nucleic acid sequence data to explore plant phylogeny has now developed into an important branch of plant systematics. [0003] Since the study of plant DNA barcodes was carried out in 2005, domestic and foreign scholars have focused on the screening of plant DNA barcodes, evaluated the applicability of each DNA candidate fragment in different groups, and finally officially proposed rbcL+matK as the standard for terrestrial plants. [0004] rbcL is the gene encoding the large subunit of...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6895
Inventor 孙子奎王锋丁方美李嫦娥方婉
Owner SHANGHAI PASSION BIOTECHNOLOGY CO LTD
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