DNA was extracted using a Stool Genomic DNA kit (Beijing ComWin Biotech Co., Beijing, China) following the instructions of the manufacturer. The DNA samples were then sent to Majorbio BioPharm Technology Co. Ltd. (Shanghai, China) for polymerase chain reaction (PCR) amplification, and Illumina high-throughput sequencing. The PCR amplification of the bacterial 16S rRNA gene amplicons (V3 + V4 regions) was performed using specific primers (338F-5′-barcode + ACTCCTACGGGAGGCAGCAG-3′; 806R-5′-GGACTACHVGGGTWTCTAAT-3′). The reaction was carried out with an initial melting step of 95°C for 3 min, followed by 27 cycles of 30 s at 95°C, 30 s at 55°C, and 45 s at 72°C, and a final elongation step of 10 min at 72°C. The 16S rDNA amplification products were electrophoresis in 2% agarose gel, and then detected and recorded on the MultiImager. The AxyPrepDNA gel recovery kit (Axygen, USA) was used to recover the PCR products. The DNA amplicons were used to conduct the high-throughput sequencing on an Illumina MiSeq platform.
Stool genomic dna kit
Manufactured by CoWin Biotech
Sourced in China
The Stool Genomic DNA kit is a laboratory tool used for the extraction and purification of genomic DNA from stool samples. It is designed to isolate high-quality DNA for downstream applications such as PCR amplification, sequencing, and other molecular biology techniques.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using stool genomic dna kit
1
Gut Microbiome DNA Sequencing Protocol
2
Fecal Microbiome Profiling with Illumina MiSeq
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Stool Genomic DNA Kit (Beijing ComWin Biotech Co., Ltd., China) was utilized to extract fecal DNA in accordance with the instructions. Amplification of the V4 region (520F: 5-AYTGGGYDTAAAGNG-3, 802R: 5-TACNVGGGTATCTAATCC-3) of 16S rRNA was carried out by PCR. The amplified DNA was purified with an AxyPrep DNA Gel Extraction Kit (Axygen Biosciences, United States), and quantified with a Quantus™ Fluorometer (Promega, United States). Purified amplicons were paired-end sequenced on an Illumina MiSeq PE300 platform in Shanghai Personalbio Co., Ltd. The resulting sequences were merged using FLASH (v1.2.11) and quality filtered using Fastp (0.19.6). Then, the high-quality sequences were denoised using the DADA2 plugin in the QIIME 2 (version 2020.2) pipeline with the suggested parameters. Using the Naive Bayes consensus taxonomy classifier implemented in QIIME 2 and the SILVA 16S rRNA database (v138), amplicon sequence variants taxonomy was determined. The Majorbio Cloud Platform's free online platform (cloud.majorbio.com) was utilized to analyze the 16S rRNA microbiome sequencing data.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!