CDNA-AFLP method suitable for gene difference expression researches of wire grass

Abstract

The invention relates to a cDNA-AFLP method suitable for gene difference expression researches of wire grass, and belongs to the field of biotechnology. The cDNA-AFLP method adopts a dedicated and mature cDNA-AFLP technical system suitable for wire grass according to the expression characteristics of the genomes of wire grass, can be used for the gene difference expression researches of wire grass, and aims to excavate specific functional genes. The cDNA-AFLP method is characterized by comprising the following steps: 1) synthesizing and purifying double-strand cDNA; 2) digesting a restriction enzyme; 3) connecting joints of the digested products; 4) conducting PCR pre-amplification on the connected product; 5) carrying out selective PCR amplification; 6) carrying out polyacrylamide gel electrophoretic analysis. According to the cDNA-AFLP method provided by the invention, no order information is required to be known in advance, the polymorphism is high, the false positivity is low, the speed is high, the suitability for comprehensive analysis of a transcriptome of wire grass is higher, and the promoting role in excavation and clone of important genes of wire grass, marker-assisted selection breeding, researches in comparative genomics and the like is important.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a cDNA-AFLP method suitable for research on differential expression of bermudagrass genes. Background technique [0002] Bermudagrass ( Cynodondactylon (L.) Pers.) is a plant of the genus Bermudagrass in the Poaceae Sansevieriae subfamily. It is an important warm-season turfgrass, forage and soil and water conservation plant. It has strong drought resistance, salt resistance, cold resistance, and trampling resistance. Few pests and diseases, fast growth, thick grass layer, widely used in warm temperate, subtropical and tropical sports lawns, parks, roads and factory greening, slope protection of rivers and hillsides, grazing grasslands, etc. It plays a very important role in the production of lawns and pastures status. [0003] Molecular markers can reflect differences in plant genetic basis at the DNA level, and are an emerging technology in the field of plant genetics and breeding....

AI Technical Summary

Problems solved by technology

[0005] However, the process of cDNA-AFLP technology is complicated, and the operation is cumbersome, and the design of primers often varies according to the plant genome. 2+ The ratio of dNTPs and dNTPs will directly affect the results of the experiment, making system development extremely difficult and limiting the application of cDNA-AFLP technology in the field of plant research (ChristianW.B.Bachem, RonaldJ.F.J.Oomen, RichardG.F.Visser, TranscriptimagingwithcDNA - AFLP: step-by-step protocol, Plant Molecular Biology Reporter, 1998, 16:157–173.)

Bermudagrass has no genome-wide information so far, and no research on differentially expressed genes for different traits using cDNA-AFLP technology

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