Feature sequences, tag primer and identification method of carya illinoensis variety Nacono
A technology of thin-shell hickory nuts and labeled primers, which is applied in the fields of botanical equipment and methods, biochemical equipment and methods, and microbial measurement/inspection, and can solve instability, low yield, inconvenient variety identification, and popularize new variety cultivation and other problems, to achieve the effect of improved reliability and simple method
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[0037] (1) Extraction of Genomic DNA of Thin-shelled Hickory Nut Variety:
[0038]Take 0.01g of the young leaves of the pecan variety to be tested, add liquid nitrogen and grind them thoroughly, and use the SDS-CTAB method to extract the genomic DNA. thing. 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. DNA samples with OD260 / OD280>1.8 were used for subsequent PCR amplification. DNA extracts were stored in a -20°C refrigerator for later use.
[0039] (2) Design specific PCR amplification primers, the sequence of the primer pair is:
[0040] The upstream primer: 5'-GTCACTTCCCTTTCTCGGCA-3'; and the downstream primer: 5'-CCCCGGGTGCAGGTATAAAA-3' were synthesized by Shanghai Bioengineering Technology Co., Ltd.
[0041] (4) PCR amplification:
[0042] Composition of PCR reaction sol...
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