Antibiotic Resistance Protocols (Methods in Molecular Biology)
139 Spike in 50 ng of the internal control Transfer the suspension to the homogenization tube and make sure it is tightly close ...
140 Add 950 μL lysing buffer supplemented with 10% 2- mercaptoethanol for the Purelink RNA mini kit (see Note 4). Continue with ...
141 Use liquid culture at late exponential phase. Prepare seven decimal dilutions of the culture in triplicate. Use one set of ...
142 successful one, otherwise it will be treated as a failed extraction that should be repeated. Extraction efficiency could be ...
143 or CT = −3.32 × Log(concentration) – intercept. Aim for the efficiency of 90–100%, i.e., E = 0.9–1.0. The efficiency can be ...
144 Incubation time can be up to 1 h. Optimization has been carried out to ensure that there is no cross-reaction between these ...
145 curve composing pure RNA with six decimal dilutions, with the highest concentration of 10 ng/μL while the lowest con- centra ...
147 Stephen H. Gillespie (ed.), Antibiotic Resistance Protocols, Methods in Molecular Biology, vol. 1736, https://doi.org/10.100 ...
148 importance of confirming morpholino findings with mutant exper- iments [ 5 ]. Recently, several methods have been establishe ...
149 suspension to solubilize the agarose and cool in a 37 °C water bath before use. Dissecting stereomicroscope with transmitte ...
150 Collect freshly laid embryos from the fish tank into a petri dish (see Note 7). Decant aquarium water from the dish and ref ...
151 Transfer 40 mL of culture into 50 mL centrifuge tube, and spin down at 5000 × g for 10 min at 4 °C. Based on OD 600 reading ...
152 Return plate to incubator and leave to “rest” for 90 min. Pour off E3 and refill with fresh E3 (see Note 18). Transfer embr ...
153 For bacterial population dynamics experiments, zebrafish embryos should be infected as normal, but care should be taken to p ...
154 tiple comparisons (e.g., a range of mutants tested versus one control parental strain). For bacterial numbers recovered fro ...
155 Volumes of 0.5–1 nL are typically used and morpholino con- centrations vary between 200 μM and 1 mM to effectively knock do ...
156 to place freshly infected embryos in a set volume of E3 (e.g., 100 μL) within 96-well plates wells. It is also advisable to ...
157 Stephen H. Gillespie (ed.), Antibiotic Resistance Protocols, Methods in Molecular Biology, vol. 1736, https://doi.org/10.100 ...
158 detection extremely challenging for clinical studies, and explains the paucity of effective therapeutic agents. These “invis ...
159 temperature (see Note 2) and add 100 mL of Albumin Dextrose Complex (ADC) (see Subheading 2.1, item 7). Middlebrook 7H11 Ag ...
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