Antibiotic Resistance Protocols (Methods in Molecular Biology)
119 Amber-colored tubes, 0.5 and 1.5 mL (Alpha Laboratories). 2. Nuclease-free water. 3. Kreatech Universal Linkage System (ULS ...
120 tion, washing the universals with GTC solution to ensure all bacilli are recovered. Spin in a single universal tube per samp ...
121 DNase I treat the samples to remove contaminating DNA, using the Qiagen RNase-free DNase kit. Apply 80 μL of DNase I: buffe ...
122 To purify the cDNA, add 250 μL cDNA binding buffer to each sample, and mix by pipetting. Transfer the samples onto the cDNA ...
123 Loosen cap ¼ turn, snap off the bottom closure and place the column into a 2 mL collection tube. Centrifuge the column at 1 ...
124 After hybridization, fill one staining trough with ~400 mL room temperature Agilent Gene Expression Wash buffer 1 (trough 1 ...
125 tion ceases on addition of GTC solution and nuclease action is minimized to stabilize the RNA signature. Solutions/centrifu- ...
126 Use an incubator for the IVT reaction or a PCR-block with variable heated-lid, so condensation does not build up on the tub ...
127 Acknowledgments L.M.W. was funded by a Brazilian government agency CAPES (Coordination for the Improvement of Higher Educati ...
128 Honeyborne I, McHugh TD, Kuittinen I, Cichonska A, Evangelopoulos D, Ronacher K, van Helden PD, Gillespie SH, Fernandez-Rey ...
129 Stephen H. Gillespie (ed.), Antibiotic Resistance Protocols, Methods in Molecular Biology, vol. 1736, https://doi.org/10.100 ...
130 that are highly supercoiled where conformational stresses cause small sections of single stranded DNA to be exposed. This re ...
131 32 P labeled dCTP (EasyTides Deoxycytidine 5′-triphosphate. (alpha^32 P), PerkinElmer, London, UK). Unincorporated nucleoti ...
132 included angle of 120° with an initial switch time of 5 s and a final switch time of 45 s at 10 °C for 22 h on a CHEF II PFG ...
133 (60 μL) is added to the top of the gel filtration column fol- lowed by 320 μL of 0.1 M Tris pH 7.5. The filtrate flows throu ...
134 4 Notes The amount of enzyme used is crucial for the success of the experiment and the activity of this enzyme varies betwe ...
135 Water used here and throughout method for all buffers is dou- ble distilled water which is then autoclaved and kept in 2 L ...
136 References Schwartz DC, Cantor CR (1984) Separation of yeast chromosome-sized DNAs by pulsed field gradient gel electrophor ...
137 Stephen H. Gillespie (ed.), Antibiotic Resistance Protocols, Methods in Molecular Biology, vol. 1736, https://doi.org/10.100 ...
138 procedure, an internal control (IC) must be included. In this assay we use a fragment of potato RNA, with known concentratio ...
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