BLOOD AGAR BASE NO.2

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Store the dehydrated medium at 10-30°C.	-pH 7.4 ± 0.2 @ 25°C -Cool to 45-50°C and add 7% sterile blood.

Upstream tips
-Store the dehydrated medium at 10-30°C.
Protocol tips
-pH 7.4 ± 0.2 @ 25°C -Cool to 45-50°C and add 7% sterile blood.

Publication protocol

H. pylori N6 wild type and tlpD mutant were tested for growth defects and behaviour in agar diffusion assays and steady-state tracking assays under iron-limited conditions by adding the iron-chelator 2,2-dipyridyl (Sigma-Aldrich, St. Louis, USA). For agar diffusion assays, bacteria grown on blood agar plates for about 20 h were resuspended in BHI medium to an OD600 of 0.07. 100 μl of the bacterial suspension were plated on Blood Agar Base No. 2 plates (Oxoid) supplemented with 5% horse serum. 10 μl of 10 mM, 20 mM and 40 mM 2,2-dipyridyl (dissolved in ethanol) as iron chelator or 10 μl of appropriate dilutions of ethanol in PBS (negative control) were applied to the plates on paper discs (diameter 5 mm). The diameter of the growth inhibition zone was measured upon incubation of bacteria at standard conditions for 48 to 72 h. Tracking assays under iron-limited conditions were performed as described above. After equilibrating the bacteria in liquid medium (see above), 0.5 mM or 5 mM 2,2-dipyridyl was added to the medium or the bacteria were left untreated. Movies were recorded 15 min and 30 min after the addition of 2,2-dipyridyl and steady-state behaviour was evaluated as described above. Steady-state assays with paraquat (10 μM, 100 μM) were carried out according to the same procedure.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Bacterial cell culture media Helicobacter pylori using BLOOD AGAR BASE NO.2 from Thermo Fisher Scientific.

Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for BLOOD AGAR BASE NO.2 below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Bacterial cell culture media Helicobacter pylori using BLOOD AGAR BASE NO.2 from Thermo Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.