Crust of the dry-aged beef were diluted with prepared sterile saline and
appropriately spread on tryptic soy agar (TSA, Difco Laboratories, USA) and
potato dextrose agar (PDA, Difco Laboratories, USA). Then, bacteria in TSA
(Difco Laboratories, USA) and yeast/mold in PDA (Difco Laboratories) were
identified using 16S rDNA and 18S rDNA sequencing, respectively (Kim et al., 2016a (link)). The chromosomal DNA of
isolated strain was separated using the BioFact Genomic DNA prep kit (BioFact,
Korea). The DNA extracts were used for the polymerase chain reaction (PCR) with
1492R (5’-GGT TAC CTT GTT ACG ACT T-3’) for bacteria and ITS4
(5’- TCC TCC GCT TAT TGA TAT GC-3’) for yeast/mold, respectively.
PCR was carried out in a LAMP-Taq programmable thermal cycler (BioFact) with one
cycle of denaturation at 95℃ for 15 min, followed by 30 cycles of
95℃ for 20 s, 50℃ for 40 s, and 72℃ for 90 s. The final
extension was carried out at 72℃ for 5 min. The purified PCR product
obtained using a PCR purification kit (BioFact) was used for a Basic Local
Alignment Search Tool (BLAST) search of sequences in the National Center for
Biotechnology Information (NCBI) database (Maidak et al., 2000 (link)).
appropriately spread on tryptic soy agar (TSA, Difco Laboratories, USA) and
potato dextrose agar (PDA, Difco Laboratories, USA). Then, bacteria in TSA
(Difco Laboratories, USA) and yeast/mold in PDA (Difco Laboratories) were
identified using 16S rDNA and 18S rDNA sequencing, respectively (Kim et al., 2016a (link)). The chromosomal DNA of
isolated strain was separated using the BioFact Genomic DNA prep kit (BioFact,
Korea). The DNA extracts were used for the polymerase chain reaction (PCR) with
1492R (5’-GGT TAC CTT GTT ACG ACT T-3’) for bacteria and ITS4
(5’- TCC TCC GCT TAT TGA TAT GC-3’) for yeast/mold, respectively.
PCR was carried out in a LAMP-Taq programmable thermal cycler (BioFact) with one
cycle of denaturation at 95℃ for 15 min, followed by 30 cycles of
95℃ for 20 s, 50℃ for 40 s, and 72℃ for 90 s. The final
extension was carried out at 72℃ for 5 min. The purified PCR product
obtained using a PCR purification kit (BioFact) was used for a Basic Local
Alignment Search Tool (BLAST) search of sequences in the National Center for
Biotechnology Information (NCBI) database (Maidak et al., 2000 (link)).
