Characteristic sequence, primers and identification method of Carya illinoinensis varieties Nacono and Sumner
A thin-shelled pecan, characteristic sequence technology, applied in biochemical equipment and methods, microbial determination/inspection, recombinant DNA technology, etc., can solve the problems of low yield, instability, cumbersome detection, etc. Sex-enhancing effect
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[0038] (1) Extraction of Genomic DNA of Thin-shelled Hickory Nut Variety:
[0039] Take 0.04g 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.
[0040] (2) Design specific PCR amplification primers, the sequence of the primer pair is:
[0041] The upstream primer: 5'-ACTGGGCCTAATCGGTTTGG-3'; and the downstream primer: 5'-CACTAGCACCCCATGAAGCA-3' were synthesized by Shanghai Bioengineering Technology Co., Ltd.
[0042] (4) PCR amplification:
[0043] Composition of PCR reaction so...
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