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Brassica juncea EST-SSR (expressed sequence tag-simple sequence repeat) marker primer group based on development of transcriptome sequence

A technology of transcriptome sequence and labeled primers, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problems of limited technology, lack, and restriction of high-quality germplasm resources, and overcome the very rare , Broaden the thinking and field, the effect of real and reliable results

Active Publication Date: 2015-08-19
ZHEJIANG UNIV
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Problems solved by technology

However, the molecular marker technology that can be applied in mustard mustard research is very limited at present. Molecular markers, as the basis of genetic linkage map and gene positioning, have become an important part of crop genetics and molecular marker-assisted breeding research and application. Therefore, the lack of mustard molecular markers The rapid and effective breeding and production application of its high-quality germplasm resources are limited
However, there is no report on the development of EST-SSR marker primers using mustard transcriptome sequences.

Method used

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  • Brassica juncea EST-SSR (expressed sequence tag-simple sequence repeat) marker primer group based on development of transcriptome sequence
  • Brassica juncea EST-SSR (expressed sequence tag-simple sequence repeat) marker primer group based on development of transcriptome sequence
  • Brassica juncea EST-SSR (expressed sequence tag-simple sequence repeat) marker primer group based on development of transcriptome sequence

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Embodiment 1

[0022] Embodiment 1: This embodiment prepares mustard mustard EST-SSR marker primer set, comprises the following steps:

[0023] (1) Acquisition of EST data of mustard mustard transcriptome:

[0024] (1.1) The young mustard leaves of one and a half month old seedlings were collected, wrapped in tin foil, frozen in liquid nitrogen, and then transferred to a ‐70°C refrigerator for storage.

[0025] (1.2) Use Trizol reagent to extract the total RNA of mustard mustard; after extracting the total RNA of the sample, use magnetic beads with Oligo (dT) to enrich mRNA; use mRNA to reverse transcribe and synthesize double-stranded cDNA, purify cDNA, and add Adenosine nucleosides were connected to the sequencing adapters, and then the 200-700bp fragments were recovered by agarose gel electrophoresis, and the recovered fragments were amplified by PCR to establish a sequencing library, using Illumina HiSeq TM In 2000, the sequencing library was sequenced; according to the sequencing resul...

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Abstract

The invention discloses a brassica juncea EST-SSR (expressed sequence tag-simple sequence repeat) marker primer group based on development of a transcriptome sequence, and belongs to the technical field of biologics. The brassica juncea EST-SSR marker primer group has the advantages that the primer group is obtained on the basis of development of the transcriptome sequence; on the basis of transcriptome sequencing, a large amount of sequence information is processed by batches, and the finding of an SSR sequence and the primer design of an SSR marker are performed, so as to overcome the defects of fewer SSR markers, low development efficiency and the like in the present brassica juncea; as the brassica juncea and brassica campestris are different materials, the effectiveness and universality of SSR primer are verified, and a foundation is laid for the development of the brassica juncea SSR primer, the auxiliary selecting of the molecule marker, the building of the genetic linkage map, the evaluation of genetic diversity, and the like.

Description

technical field [0001] The invention belongs to the field of development and application of molecular marker technology, in particular to mustard mustard EST (expressed sequence tags, ESTs) microsatellite marker primer set (EST-SSR). Background technique [0002] Mustard mustard (Brassica juncea (L.) Czern. et Coss. var. tumida Tsen et Lee) is taxonomically called stem tumor mustard, a biennial herb, and is a mustard variety produced by long-term cultivation and domestication in my country. For a long time, many researchers have done a lot of work on the collection, classification and identification of mustard germplasm resources, providing a large number of excellent materials for mustard breeding research, and breeding many mustard varieties that meet production needs. However, the molecular marker technology that can be applied in mustard mustard research is very limited at present. Molecular markers, as the basis of genetic linkage map and gene positioning, have become a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156C12Q2600/158
Inventor 陈利萍李俊星邹晓霞曹丽雯于宁宁
Owner ZHEJIANG UNIV
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