All eight bacterial strains that make up the VSL#3 product were individually provided by the manufacturer (courtesy of Actial Farmaceutical SRL, Rome, Italy), coded and cultivated as listed in S1 Table . Five ml of overnight-grown cultures were then further used for genomic DNA isolation. Cells were lyzed using lysozyme (20mg/ml), mutanolysin (10U/ml), 1% w/v SDS, 50 μg/ml RNase and 300 μg/ml proteinase K followed by an incubation of 30 min at 37 oC. Genomic DNA was then isolated from cell lysates with a RSC Blood DNA kit AS1400 according to the manufacturer’s protocol on a Promega Maxwell RCS instrument (Promega, Madison, WI, USA). DNA was quantified using Quanti-iT™ Pico Green dsDNA Assay (Invitrogen, San Diego, CA, USA).
Maxwell rcs instrument
Manufactured by Promega
Sourced in United States
The Maxwell RCS instrument is a compact and automated sample preparation system designed for the purification of nucleic acids (DNA and RNA) from a variety of sample types. The instrument utilizes magnetic bead-based technology to capture, wash, and elute nucleic acids, providing a streamlined and consistent sample preparation solution.
Automatically generated - may contain errors
2 protocols using maxwell rcs instrument
1
Genomic DNA Isolation from VSL#3 Bacteria
2
Human Genomic DNA Isolation and Purification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole blood was collected in BD Vacutainer blood collection tubes containing K 2 EDTA (BD, Franklin Lakes, NJ). Human genomic DNA (gDNA) was isolated from whole blood using the Maxwell RCS Whole Blood DNA Kit (Promega, Madison, WI) and Maxwell RCS Instrument (Promega). Two single-donor samples of anonymous human gDNA and the gDNA from seven specimens registered in the 1000 Genomes Project (NA12878, NA24385, NA24149, NA24143, NA24631, NA24694, and NA24695) were purchased from the BioChain Institute (Newark, CA) and the Coriell Institute 23, 24 (Camden, NJ), respectively.
Before library preparation, gDNA was purified by isopropanol precipitation and resuspended in 10 mmol/L Tris-HCl (pH 8.0). gDNA quality was evaluated using the BioSpec-nano (Shimadzu, Kyoto, Japan) with the following quality metrics: A260/280 Z 1.8e2.0 and A260/230 > 1.6. Furthermore, RNA contamination in the gDNA sample was assessed using the QuantiFluor One dsDNA system (Promega) and Quantus Fluorometer (Promega).
Before library preparation, gDNA was purified by isopropanol precipitation and resuspended in 10 mmol/L Tris-HCl (pH 8.0). gDNA quality was evaluated using the BioSpec-nano (Shimadzu, Kyoto, Japan) with the following quality metrics: A260/280 Z 1.8e2.0 and A260/230 > 1.6. Furthermore, RNA contamination in the gDNA sample was assessed using the QuantiFluor One dsDNA system (Promega) and Quantus Fluorometer (Promega).
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!