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Radix polygonati officinalis microsatellite DNA (deoxyribonucleic acid) molecular marker

A DNA molecule and microsatellite marker technology, applied in the direction of recombinant DNA technology, DNA/RNA fragments, etc., can solve the problems of low reproduction coefficient and few researches on molecular breeding of Polygonatum japonica

Inactive Publication Date: 2013-08-14
ANHUI NORMAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At the same time, a series of problems have appeared in the cultivation and domestication process of Polygonatum odoratum, such as low reproductive coefficient, and there are few studies on molecular breeding of Polygonatum odoratum using microsatellite markers.

Method used

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  • Radix polygonati officinalis microsatellite DNA (deoxyribonucleic acid) molecular marker
  • Radix polygonati officinalis microsatellite DNA (deoxyribonucleic acid) molecular marker
  • Radix polygonati officinalis microsatellite DNA (deoxyribonucleic acid) molecular marker

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Embodiment 1

[0024] Construction of Polygonatum Polygonatum Microsatellite DNA Enrichment Library

[0025] 1) Genome extraction and digestion:

[0026] Using the CTAB method introduced by Doyle J (Doyle J. DNA protocols for plants-CTAB total DNA isolation [A]. In Hewitt GM, Johnston A (eds.), Molecular Techniques in Taxonomy [M]. Berlin: Springer, 1991, 283 -293) to extract the genome. The genome was digested with restriction endonuclease Sau 3AI. After 2% agarose gel electrophoresis, the DNA fragments of 300-900 bp were excised under ultraviolet light. Recover with gel recovery kit.

[0027] 2) Add connectors:

[0028] Add the gel recovered and purified product to the adapter:

[0029] oligo A (5'-GGCCAGAGACCCCCAAGCTTCG-3')

[0030] oligo B (5’-PO 4 -GATCCGAAGCTTGGGGTCTCTGGCC-3'),

[0031] Under the action of T4 DNA Ligase, they were ligated overnight in a water bath at 16°C.

[0032] 3) PCR detection adapter connection:

[0033] Use oligo A as primer and ligation product as tem...

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Abstract

The invention discloses a radix polygonati officinalis microsatellite DNA molecular marker, comprising the steps of constructing a radix polygonati officinalis microsatellite (CT) n enrichment library; screening and sequencing microsatellite sequence positive clone; obtaining 55 clones containing the microsatellite repeated sequence; and screening to obtain 9 high polymorphic microsatellite molecular markers Po1, Po2, Po3, Po4, Po5, Po6, Po7, Po8 and Po9. The radix polygonati officinalis microsatellite DNA molecular marker can be used for researching genetic diversity of the radix polygonati officinalis, variation and evolutionary relationship between polygonatum plants and population, has good repeatability and is a reliable and efficient molecular marker.

Description

technical field [0001] The invention relates to a molecular marker technology, in particular to a microsatellite molecular genetic marker of polygonatum polygonatum. Background technique [0002] Microsatellites are also called short tandem repeats (Short Tandem Repeats, STRs), simple sequence repeats (Simple Sequence Repeat, SSRs), simple sequence length polymorphism (SSLP). It refers to a kind of simple tandem repeat consisting of several nucleotide pairs (called core sequence, generally 1-6 bps) as the repeating unit throughout the biological genome. The number of repeats is highly variable among different genotypes of the same species, but the two ends of this repeat are relatively conserved single-copy sequences. Based on this, primers can be designed to analyze the variation of the repeat number of the core sequence by PCR , polymorphisms were detected among different genotypes. According to the composition of repeating units, microsatellite DNA sequences are divided...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 朱国萍高鹏王晖郑基阳李雪王文才唐王刚赵晓宇
Owner ANHUI NORMAL UNIV
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