SSR molecular marker method of brassica allohexaploid and primers thereof
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A molecular marker and Brassica technology, applied in biochemical equipment and methods, microbiological determination/testing, DNA/RNA fragments, etc., can solve the problem of molecular markers and genetic maps of new allohexaploid Brassica plants Construct issues that have not been reported yet, to achieve the effects of easy observation and recording, clear bands, and reduced concentration
Active Publication Date: 2014-09-24
ZHEJIANG UNIV
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[0005] So far, although the molecular markers and the construction of genetic maps of Brassica diploid and tetraploid species have been reported, the molecular markers and genetic map construction of new allohexaploid Brassica plants have not been reported.
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[0049] The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.
[0050] 1. Preparation of main reagents
[0051] (1) DNA extraction reagent formula
[0052] ①0.5mol / L EDTA (pH=8): Weigh Na 2 EDTA·2H 2 O 186.1g, add 800mL of deionized water, place it on a magnetic stirrer and stir fully, adjust the pH to 8.0 (about 20g NaOH) with NaOH, when the pH is close to 8.0, EDTA can be completely dissolved, add deionized water to dilute to 1L, aliquoted and autoclaved, stored at room temperature.
[0053] ②1mol / L Tris–HCL (pH=8.0): Dissolve 121.1g Tris in 800mL of water, after the solution is cooled to room temperature, add 42mL of concentrated HCl to adjust the pH to 8, add water to make up to 1L, aliquot and autoclave.
[0054] ③DTT: Dissolve 5g of DTT in 32.4mL of water, aliquot and store in -20℃ refrigerator.
[0055] ④CTAB extraction buffer: Measure 10mL of 1mol / L Tris-HCl, 4mL of 0.5mol / L EDTA, wei...
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Abstract
The invention discloses an SSR molecular marker method of a brassica allohexaploid and primers thereof. The SSR molecular marker method of the brassica allohexaploid comprises the steps: hybridizing a brassica allohexaploid parent, and performing microspore culture on the obtained filial generation to obtain a DH colony; extracting genome DNAs of the brassica allohexaploid parent and the DH colony, and performing PCR amplification with all genome DNAs as a template by using the primers; constructing a genetic map of the brassica allohexaploid parent according to a PCR amplification result, wherein 217 pairs of primers are included, with base sequences respectively shown in SEQ ID No. 1/2-433/434. According to the SSR molecular marker method, a first genetic map of the brassica allohexaploid is constructed, and the basis is provided for further performing QTL location and molecular marker assistant breeding.
Description
technical field [0001] The invention relates to plant molecular marker technology, in particular to a method for SSR molecular marker of Brassica allohexaploid and primers thereof. Background technique [0002] Brassica spp. is the most important genus in Brassicaceae (formerly known as Cruciferae). In my country, Brassica crops are widely planted, including many important vegetables, oil plants, medicinal and feed crops. According to Yu's triangle (U N. Genomic analysis in Brassica with special reference to the experimental formation of B. napus and peculiar mode of fertilization [J]. The Journal of Japanese Botany, 1935, 7:389-452), the three different Source tetraploid [Brassica napus (B.napus, 2n=38, AACC), Brassica napus (B.juncea, 2n=34, AABB) and Ethiopian mustard (B.carinata, 2n=36, BBCC)] respectively It is derived from three diploid species [Brassica napus (B.rapa, 2n=20, AA), cabbage (B.oleracea, 2n=18, CC), black mustard (B.nigra, 2n=16, BB )] Genome. After p...
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