Hi !
I am pretending to do qPCR of 16sRNA of my samples, I want to watch the changes of density of bacteria through my experiment, but this is my first time doing that technique.
Could you give me some tips about how to do it ?
Greetings.
Following article may be useful for you.
Article Use of 16S rRNA Gene-Targeted Group-Specific Primers for Rea...
The primers are 341F/534R (Cf 1uM), pcr product 194, Tm 60 degrees. Thermoprofile: 16S rRNA: 15 min at 95°C of initial denaturation, 35 cycles with 15 s at 95°C, 30 s at 60°C, 30 s at 72°C and 30 s at 80°C, plus a melt curve stage with 15 s at 95°C, 15 s at 78°C and 15s at 95°C.
G. Muyzer, E. C. De Wall, and A. G. Uitterlinden, “Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reactionamplified genes coding for 16S rRNA,” Apply Environ. Microbiol., vol. 59, no. 3, pp. 695–700, 1993.
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