Stool samples collected from the hospitalized children were immediately transferred to the laboratories, and were kept in 2-8°C. The DNA was extracted from the stool samples using the QIAamp DNA Stool Mini Kit (50) (Qiagen, Germantown, MD, USA). An in-house multiplex PCR assay targeting virulence genes, which were specific for each of the five pathotypes (EAEC, enteroinvasive E. coli [EIEC], EPEC, ETEC, STEC), was performed in accordance with the method introduced by Antikainen et al. [13 (link)]. In addition, an in-house PCR assay targeting the invA gene specific for Salmonella spp. was also performed in the same stool sample according to the method reported by Cunningham et al. [14 (link)]. Table 1 describes the target genes and sequences of primers for the PCR assay. The PCR assay was considered the goldstandard to diagnose E. coli and Salmonella infections in the present study.
Qiaamp dna stool mini kit 50
Manufactured by Qiagen
Sourced in United States
The QIAamp DNA Stool Mini Kit (50) is a laboratory product designed for the purification of genomic DNA from stool samples. It provides a standardized protocol for the extraction and isolation of DNA from fecal material.
Automatically generated - may contain errors
Lab products found in correlation
Nanodrop 2000, by Thermo Fisher Scientific (1 mentions)
Nano drop spectrophotometer system nd 1000, by Thermo Fisher Scientific (1 mentions)
Ultraclean fecal dna isolation kit, by Qiagen (1 mentions)
Sherlock ax 100 isolations, by A&A Biotechnology (1 mentions)
Zr fecal dna miniprep, by Zymo Research (1 mentions)
4 protocols using qiaamp dna stool mini kit 50
1
Multiplex PCR Diagnostics for Enteric Pathogens
2
DNA Extraction from Aphid Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The collected adult aphids were surface sterilized with absolute ethanol. Then, we used sterile water to rinse off the remaining ethanol. Finally, aphids were air dried on a clean bench. After air drying, 2 mg of adults (≤ 20 adult aphids) was taken from each replicate for DNA extraction. DNA extractions were performed using a QIAamp DNA Stool Mini Kit (50) (QIAGEN, Germany) according to the manufacturer’s instructions. The purity and concentration of DNA were confirmed using 1% agarose gel electrophoresis and an ultraviolet–visible spectrophotometer (NanoDrop 2000, Thermo Scientific).
3
Fecal DNA Extraction and Quality Assessment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bacterial DNA was extracted from fecal samples using QIAamp® DNA stool Mini kit (50) (Qiagen, Inc., Valencia, CA, USA), following the manufacturer's instructions. Fecal DNA was quantified by Nano-Drop spectrophotometer system ND-1000 (Thermo Fisher Scientific Inc., Wilmington, USA). DNA quality was assessed by determining the ratio of absorbance at 260 and 280 nm. Only DNA samples that had 260:280 nm ratios higher than 1.8 were used for further analysis.
4
Comparison of Fecal DNA Isolation Methods
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fecal samples were obtained from a clinically healthy cat (confirmed by real-time PCR according to Frey et al. [27 (link)]). To compare the DNA isolation methods, samples with the following weights of feces were prepared: 145 mg (ZR Fecal DNA MiniPrep™, Zymo Research, Freiburg, Germany), 245 mg (UltraClean® Fecal DNA Isolation Kit, MO BIO Laboratories Inc., Carlsbad, CA, USA), 215 mg (QIAamp® DNA Stool Mini Kit (50), Qiagen, Hilden, Germany), and 15 mg (Sherlock AX/100 isolations, A&A Biotechnology, Gdynia, Poland). In order to maintain the weight recommended by the manufacturer, we lowered the weight of the fecal samples by 5 mg.
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!