Further culture-independent characterization of the lactic microbiota of Serro artisanal cheese

The assessment of bacterial diversity in CO2 treated and untreated raw milk during cold storage using high‐throughput sequencing technology

“…The 16S rDNA V3–V4 region of the ribosomal RNA gene was amplified through PCR (Biçer et al ., 2021; Dimov, 2022). The PCR reaction involved cycles of 94 °C for 2 min, followed by 30 cycles at 98 °C for 10 s, 62 °C for 30 s and 68 °C for 30 s, with a final extension at 68 °C for 5 min using primers 341F: CCTACGGGNGGCWGCAG; 806R: GGACTACHVGGGTATCTAAT (Witzke et al ., 2020; Shaffer et al ., 2021; Ferreira et al ., 2022). The PCR reactions were performed in triplicate, with each mixture containing 50 μL and consisting of 5 μL of 10 × KOD Buffer, 5 μL of 2 mM dNTPs, 3 μL of 25 mM MgSO 4 , 1.5 μL of each primer (10 μM), 1 μL of KOD polymerase and 100 ng of template DNA.…”

“…CCTACGGGNGGCWGCAG; 806R: GGACTACHV GGGTATCTAAT (Witzke et al, 2020;Shaffer et al, 2021;Ferreira et al, 2022). The PCR reactions were performed in triplicate, with each mixture containing 50 lL and consisting of 5 lL of 10 9 KOD Buffer, 5 lL of 2 mM dNTPs, 3 lL of 25 mM MgSO 4 , 1.5 lL of each primer (10 lM), 1 lL of KOD polymerase and 100 ng of template DNA.…”