Overnight cultures in GM17 of E. faecalis were adjusted to OD600 of 0.2 in fresh M17 supplemented with 2% glucose. One hundred microliters of the bacterial suspension were inoculated into CytoOne polystyrene microwells plate coated with 1 μg/ml of hyaluronic acid (Sigma-Aldrich), chondroitin sulfate (Carl Roth, Karlsruhe, Germany), or heparin sodium (ThermoFisher). After 24 or 48 h of incubation at 37°C, the plates were washed with 0.9% NaCl to remove unbound bacteria. Each well was then stained with 0.1% (wt/vol) crystal violet for 15 min at room temperature. Wells were then rinsed two times with 0.9% NaCl. Adherent cells were dissolved in 30% acetic acid, and the OD550 was measured using a microplate reader Nano Tecan (Life Science). At least three experiments were performed for each condition.
Chondroitin sulfate
Manufactured by Carl Roth
Sourced in Germany
Chondroitin sulfate is a natural and complex polysaccharide found in the cartilage and connective tissues of the human body. It is commonly used in the production of various laboratory and research materials.
Automatically generated - may contain errors
Lab products found in correlation
Sodium heparin, by Thermo Fisher Scientific (1 mentions)
Hyaluronic acid, by Merck Group (1 mentions)
Ethylene glycol tetraacetic acid (egta), by Thermo Fisher Scientific (1 mentions)
Ethylene glycol tetraacetic acid (egta), by Carl Roth (1 mentions)
β mercaptoethanol, by Thermo Fisher Scientific (1 mentions)
2 protocols using chondroitin sulfate
1
Biofilm Formation of E. faecalis
2
Assessing DNA Origami Stability in Cell Media
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To assess the stability
of the DNA
origami in cell medium supplemented with FBS (Figure S1 ), bare and coated disks were incubated in cell medium
with 10% FBS (PAN-Biotech) at 37 °C for 2 h in a total volume
of 10 μL. The incubation was stopped through deactivating the
FBS nucleases by adding β-mercaptoethanol (1.2 μL, Thermo
Fisher) and EGTA (75 mM, 0.8 μL, Thermo Fisher) and incubating
at 37 °C for 30 min (final concentration 5 mM EGTA and 10% β-mercaptoethanol).
To verify the integrity of the DNA origami disk, the coating was removed
to allow the sample to run on AGE: 1 μL of chondroitin sulfate
(0.5 M, Carl Roth) was added, and the concentration of MgCl2 was adjusted to restore a concentration of 20 mM. The samples were
incubated at 37 °C for 1 h to remove the coating and successively
loaded on agarose gel (2% agarose, 1X TBE, 15 mM MgCl2,
1X SybrSafe, run at 70 V for 90 min in an ice-water bath).
of the DNA
origami in cell medium supplemented with FBS (
with 10% FBS (PAN-Biotech) at 37 °C for 2 h in a total volume
of 10 μL. The incubation was stopped through deactivating the
FBS nucleases by adding β-mercaptoethanol (1.2 μL, Thermo
Fisher) and EGTA (75 mM, 0.8 μL, Thermo Fisher) and incubating
at 37 °C for 30 min (final concentration 5 mM EGTA and 10% β-mercaptoethanol).
To verify the integrity of the DNA origami disk, the coating was removed
to allow the sample to run on AGE: 1 μL of chondroitin sulfate
(0.5 M, Carl Roth) was added, and the concentration of MgCl2 was adjusted to restore a concentration of 20 mM. The samples were
incubated at 37 °C for 1 h to remove the coating and successively
loaded on agarose gel (2% agarose, 1X TBE, 15 mM MgCl2,
1X SybrSafe, run at 70 V for 90 min in an ice-water bath).
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!