By Stephen H. Gillespie
Scientific and diagnostic microbiologists aspect molecular and actual tools for learning the transforming into challenge of antibiotic resistance in micro organism, and facilitate new antibiotic learn courses to assist redress the matter. The ideas diversity from those who supply speedy prognosis via DNA amplifications and phage reveal, 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 advance and enforce now diagnostic and epidemiological instruments.
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Additional resources for Antibiotic Resistance: Methods and Protocols
And Jacobs, W. , Jr. (1997) Conditionally replicating luciferase reporter phages: improved sensitivity for rapid detection and assessment of drug susceptibility of Mycobacterium tuberculosis. J. Clin. Microbiol. 35, 3232–3239. 7. Riska, P. , Jacobs, W. , Bloom, B. , and Chan, J. (1997) Specific identification of Mycobacterium tuberculosis with the luciferase reporter mycobacteriophage: use of p-Nitro-a-Acetylamino-b-Hydroxy propiophenone. J. Clin. Microbiol. 35, 3225–3231. 8. , and Drobniewski, F.
Use Mtb-specific PCR MIMIC stock solution (100 attomoles/µL) to prepare eight 10-fold serial dilutions of the PCR MIMIC (M1 through M8, 10 through 10–6 attomoles/µL, respectively), using MIMIC Dilution solution as diluent. The dilution series can be stored at –20°C and discarded after three uses. 2. 4 µL. Multiply the amount of each ingredient by the total number of the QSTNPCR reactions, combine the solutions, and mix them gently by pipeting. Aliquot 48 µL of the master mix to each labeled PCR tube.
3. SECONDARY PCR AMPLIFICATION FOR THE PRODUCTION OF MTB-SPECIFIC PCR MIMIC 1. Dilute 2 µL of the primary PCR product to 200 µL in H2O. 2. 6 µL. Thus, the final reaction volume for this PCR is 100 µL. 3. Overlay the reaction mixture with 100 µL light mineral oil. 4. Carry out 25 PCR cycles, using the same cycling parameters described above for the primary PCR. 5. Electrophorese 5 µL of the secondary PCR product on an 1% agarose gel. Again, a 486-bp band should be observed. 4. PURIFICATION OF THE MTB-SPECIFIC PCR MIMIC 1.
Antibiotic Resistance: Methods and Protocols by Stephen H. Gillespie