Received: 01-03-2023
Accepted: 05-10-2023
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Identification of the Mating-type Loci of Cordyceps militarisby Multiplex PCR
,
Vo Ba Duy
1
,
Dinh Xuan Tu
2, 3
Keywords
Cordyceps militaris, molecular marker, MAT, multiplex PCR
Abstract
In the cross-breeding of Cordyceps militaris using single spores, determining the genotype of the mating type locus is crucial for constructing a cross diagram. Individual PCRshave been conducted to identify the mating type loci MAT1-1 and MAT1-2 in this fungus. The research aimed to design specific primers for Multiplex PCR to efficiently and simply identify mating-type loci in a shorter time. The primer sequences were derived from the sequences of two mating type loci MAT1-1 (AB194982) and MAT1-2 (AB084257) retrieved from the GenBank database using the Primer-BLAST tool. The research results indicate that the Multiplex PCR, comprising three primer pairs MAT1-1-1-MUL, MAT1-1-2-MUL, and MAT1-2-1-MUL, successfully amplified products with sizes of 527bp, 323bp, and 222bp from the MAT1-1-1, MAT1-1-2, and MAT1-2-1 genes, respectively. The Multiplex reaction exhibits high specificity and reliability in determining mating genotypes in C. militaris, making it a valuable tool for applications in breeding selection based on molecular markers in C. militaris.
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