By Stephen H. Gillespie
Scientific and diagnostic microbiologists element molecular and actual equipment for learning the growing to be challenge of antibiotic resistance in micro organism, and facilitate new antibiotic learn courses to assist redress the matter. The suggestions diversity from those who supply quick analysis via DNA amplifications and phage exhibit, to these for plotting the transmission of resistant organisms and investigating their epidemiology. in addition to illuminating the fundamental biology of antimicrobial resistance, additionally they boost and enforce now diagnostic and epidemiological instruments.
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Additional info for Antibiotic Resistance: Methods and Protocols
7. To calculate the amount of the 603-bp qX174 DNA fragment, use the following equation: ng of qX174 DNA fragment = (size of qX174 DNA fragment) (ng of qX174 DNA)/(sum of all qX174 DNA fragments). 19 ng. 78 ng of DNA, respectively. 8. 4. under Densitometry for details). 9. Convert nanograms to molar quantities and calculate the concentration of the Mtb-specific PCR MIMIC in attomoles/µL (see Note 5). 10. Dilute the concentrated Mtb-specific PCR MIMIC solution to 100 attomoles/µL with the MIMIC Dilution Solution.
DNA Thermal Cycler. 32. 100 bp DNA Ladder. 33. Model GS-700 Imaging Desitometer (Bio-Rad, Hercules, CA). 34. 18% sodium sulfite solution. Store solution in a well-capped brown bottle. 35. Polaroid® type 667 B&W film. 36. Polaroid® type 665 positive/negative film. 37. 625 mg/mL solution in a dropper bottle. Shield the solution from light. 5 µg/mL. Store at room temperature. 38. Agarose I (Amresco). 39. Agarose 3:1, High Resolution Blend (Midwest Scientific, St. Louis, MO). 40. Electrophoresis buffer: Prepare 5X Tris-borate buffer (TBE) stock solution and store at room temperature.
Plastic trays large enough to immerse the electrophoresis plate. 2. , Luckham R100/TW Rotatest shaker). 3. Fix/stop solution 10% acetic acid: 200 mL glacial acetic acid in 1 800 mL deionized water. 4. Staining solution: combine 2 g AgNO3 with 3 mL 37% formaldehyde in 2 L deionized water. 5. Developing solution: dissolve 60 g Na2CO3 in 2 L deionized water. Chill to 10°C. Immediately before use add 3 mL 37% formaldehyde and 400 µL sodium thiosulphate (10 mg/mL). 3. 1. Organisms and DNA Extraction M.
Antibiotic Resistance: Methods and Protocols by Stephen H. Gillespie