Primerscript pcr kit perfect real time

Bacterial DNA in the ileal and cecal digesta was extracted by using the Stool DNA Kit (Omega Bio-tek) according to the manufacturer’s instruction. The microbial real-time quantitative PCR was determined as described previously [34 (link)]. Briefly, the number of total bacteria was analyzed by real-time quantitative PCR using SYBR Premix Ex Taq reagents (TaKaRa Biotechnology (Dalian) Co., Ltd., Dalian, China) and CFX-96 Real-Time PCR Detection System (Bio-Rad Laboratories, Richmond, CA), and the number of Lactobacillus, E. coli and Bifidobacterium was analyzed by real-time quantitative PCR using PrimerScriptTM PCR kit (Perfect Real Time; TaKaRa Biotechnology (Dalian) Co., Ltd., Dalian, China) and CFX-96 Real-Time PCR Detection System (Bio-Rad Laboratories, Richmond, CA) as previously described (Chen et al., 2013). All primers and probes were purchased by TaKaRa Biotechnology (Dalian) Co., Ltd. (Dalian, China), which was listed in Table 3. For the quantification of bacteria in the test samples, specific standard curves were generated by constructing standard plasmids as presented by Chen et al. (2013) [34 (link)]. In addition, bacterial copies were transformed (log₁₀) before statistical analysis.

Bacterial DNA in jejunal and cecal digesta were extracted by using the Stool DNA Kit (Omega Bio-tek) according to the manufacturer's instruction. The microbial real-time quantitative PCR was determined as described previously [30 (link)]. Briefly, the number of total bacteria was analyzed by real-time quantitative PCR using SYBR Premix Ex Taq reagents (TaKaRa Biotechnology (Dalian) Co, Ltd., Dalian, China) and CFX-96 Real-Time PCR Detection System (BioRad Laboratories, Richmond, CA), and the numbers of Bacillus, Lactobacillus, E. coli, and Bifidobacterium were analyzed by real-time quantitative PCR using PrimerScript TM PCR kit (Perfect Real-Time; TaKaRa Biotechnology (Dalian) Co, Ltd, Dalian, China) and CFX-96 Real-Time PCR Detection System (Bio-Rad Laboratories, Richmond, CA) as previously described [30 (link)]. All primers and probes (Table 3) were purchased from TaKaRa Biotechnology (Dalian) Co, Ltd. (Dalian, China). For the quantification of bacteria in the test samples, specific standard curves were generated by constructing standard plasmids as presented by Chen et al [30 (link)]. In addition, bacterial copies were transformed (log₁₀) before statistical analysis.

According to the manufacturer's instruction, bacterial DNA in cecal digesta was extracted with the Stool DNA Kit (Omega Bio-tek, Doraville, GA). The microbial real-time quantitative PCR was determined as described previously [11 (link)]. Briefly, the number of total bacteria was analyzed by real-time quantitative PCR using SYBR Premix Ex Taq reagents (TaKaRa Biotechnology (Dalian) Co., Ltd., Dalian, China) and CFX-96 Real-Time PCR Detection System (Bio-Rad Laboratories, Richmond, CA), and the number of Lactobacillus, E. coli and Bifidobacterium was analyzed by real-time quantitative PCR using PrimerScript^TM PCR kit (Perfect Real Time; TaKaRa Biotechnology (Dalian) Co., Ltd., Dalian, China) and CFX-96 Real-Time PCR Detection System (Bio-Rad Laboratories, Richmond, CA). All primers and probes, listed in Supplementary Table 2, were purchased by TaKaRa Biotechnology (Dalian) Co., Ltd. (Dalian, China). For the quantification of bacteria in test samples, specific standard curves were generated by constructing standard plasmids. Bacterial copies were transformed (log₁₀) before statistical analysis.