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
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[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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