Mustard est‑ssr marker primer set developed based on 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
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[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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