Bacterial DNA was isolated from jejunal content using stool DNA extraction kit (Omega Bio-Tek, Doraville, CA, USA) following the manufacturer’s instructions. The purity and concentration of DNA in each sample were measured by Nanodrop 2000. The populations of total bacteria, Lactobacillus, Enterococcus, Salmonella, Bifidobacterium, Escherichia coli, Enterobacter, and Clostridium were determined by q-PCR as described by Rezaei et al. (2015) [32 (link)]. The primers used for q-PCR were obtained from published works [32 (link)] and are listed in Table S3 . Standard curves were prepared with serial dilutions of PCR products from pure cultures of the different bacterial groups, and copies per sample were calculated using Ct values and standard curves.
Stool dna extraction kit
Manufactured by Omega Bio-Tek
Sourced in United States
The Stool DNA Extraction Kit by Omega Bio-Tek is a laboratory tool designed to isolate and purify DNA from human stool samples. The kit provides a standardized process for extracting high-quality DNA, which can be used for various downstream applications such as genetic analysis and research.
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Lab products found in correlation
2 protocols using stool dna extraction kit
1
Quantifying Gut Microbial Populations
2
Quantification of Gut Microbiome Composition
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Bacterial DNA from the caecal and colonic digesta were isolated using the stool DNA extraction kit (Omega Bio-tek, Doraville, CA, USA) according to the manufacturer's instructions. Primers and probes for total bacteria, Escherichia coli, Lactobacillus and Bifidobacterium used in this experiment are presented in Table S2† and designed according to the latest report,19 (link) and commercially synthesized by Life Technologies Limited. Briefly, the quantification of total bacteria was conducted by using 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), and the quantification of Escherichia coli, Lactobacillus and Bifidobacterium were carried out by real-time quantitative PCR using PrimeScript™ PCR kit (Perfect Real Time; Takara) and CFX-96 Real-Time PCR Detection System (Bio-Rad Laboratories) as previously described.20,21 (link) Standard curves were manufactured according to the previous study.21 (link) All reactions were specific for the target species. The quantitative PCR data were normalized based on the DNA concentrations. Bacterial copies were transformed (log 10) before statistical analysis.
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