The bacterial density of AKM and LGG in the fecal samples was estimated using quantitative real-time polymerase-chain reaction (qPCR) with species specific primers (AKM_Fwd: 5’-CCT TGC GGT TGG CTT CAG AT-3’ and AKM_Rev: 5’-CAG CAC GTG AAG GTG GGG AC-3’85 (link) and LGG_Fwd: 5’-GCC GAT CGT TGA CGT TAG TTG G-3’ and LGG_Rev: 5’-CAG CGG TTA TGC GAT GCG AAT-3’86 (link)) purchased from Integrated DNA Technologies. Standard curves (Table S2) were based on a dilution series of total DNA extracted from monocultures of AKM and LGG. The qPCR results were obtained using the CFX96 Touch Real-Time PCR Detections System (Bio-Rad Laboratories) and the reagent SsoFast EvaGreen Supermix with Low ROX (Bio-Rad Laboratories, cat. No. 1725211), and run as previously described87 (link).
Protocol detail
Quantitative Analysis of Gut Microbiome
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization : University of Copenhagen
Top 5 similar protocols
Variable analysis
independent variables
- Bacterial density of AKM and LGG in the fecal samples
- Bacterial density of AKM and LGG in the fecal samples measured using qPCR
- Species-specific primers for AKM and LGG
- Standard curves based on a dilution series of total DNA extracted from monocultures of AKM and LGG
- CFX96 Touch Real-Time PCR Detections System (Bio-Rad Laboratories)
- SsoFast EvaGreen Supermix with Low ROX reagent (Bio-Rad Laboratories)
- Positive control: Monocultures of AKM and LGG used to generate standard curves
- Negative control: Not explicitly mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
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!