Molecular distinctness labeling primers for aster dendrobium and detection method
A technology for labeling primers and specificity, which is used in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. It can solve the problems of long cycle, poor repeatability and low specificity, and achieve the effect of simple method.
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[0037] (1) Extraction of genomic DNA of Dendrobium species: Take 0.01g of young leaves of Dendrobium species to be tested, add liquid nitrogen to grind thoroughly, and use SDS-CTAB method to extract genomic DNA. After repeated extractions, extract the genome of Dendrobium Crude DNA extract. The crude DNA was purified by Magabio Nucleic Acid Purification Kit (Bioer, Hangzhou, China), and detected by 1.5% agarose gel electrophoresis and DNA / RNA UV spectrophotometer (GeneQuant Pro, GE Healthcare). Integrity, Purity and Concentration. OD 260 / OD 280 DNA samples >1.8 were used for subsequent PCR amplification. DNA extracts were stored in a -20°C refrigerator for later use.
[0038] (2) Design specific PCR amplification primers. The sequence of the primer pair is: 5'-AGCGGCCGCGCCTTCC-3' and the downstream primer 5'-GGAAGGCGCGGCCGCT-3', synthesized by Hangzhou Qingke Engineering Technology Co., Ltd.
[0039] (3) PCR amplification of SCAR molecular markers:
[0040] The composit...
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