By Bill J Mitchell
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1). The PCR efficiency of the RNA standards should be between 90 and 100%. The controls should be reading N/A (no amplification) or at an acceptable threshold cycle (CT) level. Create data reports which can be generated by the iQ software. Export the data into an excel sheet to analyze data. 4. A standard curve is necessary in order to quantify your unknown RNA sample. Since RNA is being used in these assays instead of DNA, a RNA standard curve must be generated and run with each plate of unknown samples.
Bull WHO 52, 429–436. 18. A. (1978) Interrelationships among arenaviruses measured by indirect immunofluorescence. Intervirology 9, 344–350. 19. L. (1979) Immunofluorescent staining. H. J. Ed. Diagnostic Procedures for Viral, Rickettsia, and Chlamydial Infections, 5th Edn. American Public Health Association, Washington, DC. Chapter 3 Detection of Human Caliciviruses in Fecal Samples by RT-PCR Robert L. Atmar, Frederick H. Neill, and Françoise S. Le Guyader Abstract Human caliciviruses, noroviruses in particular, are a common cause of gastroenteritis in persons of all age groups.
7. Bio-Rad iQ4 real-time detection system or equivalent (Hercules, CA). Quantitect probe reverse transcriptase polymerase chain reaction (RT-PCR) kit (Qiagen no. 204445) or equivalent. Optical plates in a 96-well or 48-well format (Bio-Rad no. 2239441) with sealing tape (Bio-Rad no. 2239444). SYBR green super mix kit for optimization step (Bio-Rad no. 170-8880). Various consumable (tips, 15 ml tubes, water, etc). Optimized sequence specific oligonucleotides (primers and probes) see Table 2.
Advanced oil well drilling engineering: Handbook and computer programs by Bill J Mitchell