Chinese cabbage est‑ssr marker and its primers and applications
A Chinese cabbage, labeling technology, applied in the field of molecular biology, can solve the problem that the Chinese cabbage EST-SSR primer is not saturated
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Embodiment 1
[0038] Example 1. Development of EST-SSR primers
[0039] The EST sequences used to develop EST-SSR primers came from the high-throughput transcriptome sequencing of Chinese cabbage (B.rapassp.pekinensis) rosette leaves and ball leaves by our research group (Wang et al., 2012). Through transcriptome sequencing, a total of 53.47 million high-quality EST sequences were obtained. Trinity software was used for these EST sequences ( http: / / trinityrnaseq.sourceforge.net / ) were assembled to obtain the corresponding contig and unigene, and then the TGICL software package (TIGR Gene Indices Clustering) was used to remove redundancy, and a total of 52,000 unigenes were obtained. In this study, the MISA software (http: / / www.pgrc.ipk-gatersleben.de / misa) was used to analyze and identify the EST-SSR of these 52,000 Unigenes (standards refer to Zhang et al., 2004), and a total of 10,400 EST-SSRs. Through the Chinese cabbage SSR database ( http: / / oilcrops.info / SSRdb ), a total of 27...
Embodiment 2
[0043] Application of embodiment 2.EST-SSR labeled primers
[0044] 2.1 Extraction of plant material and template DNA
[0045] Select 24 varieties (lines) of Chinese cabbage: 682, Guangdong Zao, Zaohuangbai, Z61-8, Fushan Baotou, Li-3, 212-7, Tianjin Qingmaye, Kuaicai No. 6, Golden Cabbage J, Siji Chinese cabbage, Siji Huangyang Xiaobaicai, Pinzao No. 1, Hanyu's special selection of yellow heart, Almighty Four Seasons Express, Jingyou Xiaobaicai Express, Korean Golden Baby, Keyi Xia Baby, Jinnuo Chunqiu Baby Cai, Four Seasons Green Stem small bitter vegetables, Oily Green 157, Tuyao rape, Degao Youliang green stems, Qingxiu F1 green stems. Marked as 1-24 groups respectively.
[0046] The total DNA was extracted by the modified CTAB method, and the specific steps were as follows:
[0047] 1. Take 1-10g of fresh Chinese cabbage leaves and quickly grind them into powder in liquid nitrogen;
[0048] 2. Transfer the frozen powder into a pre-cooled centrifuge tube, add an equal ...
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