With RFLP analysis, one utilizes homologous DNA differences within restriction enzyme binding sites. By fragmenting DNA samples into pieces, agarose gel electrophoresis may be carried out to separate the fragments based on length and transferred to a membrane via Southern blot followed by probe hybridization. RFLP analysis is often used for DNA profiling.
Real-time PCR on the other hand, utilizes heat-stable DNA polymerases to amplify minute quantities of DNA especially when more DNA template is required for further reactions/analysis. By prompting the reaction to occur by adding other components such as MgCl2 buffers, dNTPs, and dH2O in an appropriate qPCR machine, one may view the melting curve and amplification process real-time.
Sometimes, PCR-RFLP may be carried out to amplify insufficient DNA templates through PCR at a significantly shorter time before subsequently carrying out RFLP analysis.
With RFLP analysis, one utilizes homologous DNA differences within restriction enzyme binding sites. By fragmenting DNA samples into pieces, agarose gel electrophoresis may be carried out to separate the fragments based on length and transferred to a membrane via Southern blot followed by probe hybridization. RFLP analysis is often used for DNA profiling.
Real-time PCR on the other hand, utilizes heat-stable DNA polymerases to amplify minute quantities of DNA especially when more DNA template is required for further reactions/analysis. By prompting the reaction to occur by adding other components such as MgCl2 buffers, dNTPs, and dH2O in an appropriate qPCR machine, one may view the melting curve and amplification process real-time.
Sometimes, PCR-RFLP may be carried out to amplify insufficient DNA templates through PCR at a significantly shorter time before subsequently carrying out RFLP analysis.
Both are two different techniques. RFLP allows to identify DNA fragments based on unique patterns of restriction enzyme cutting in specific regions of DNA and see them in gel.whereas, Real time PCR, is an amplification of your target gene using specific primers and you can monitor the reaction in real time.
Hi there,
RFLP allows to differentiate DNA according to their content in restriction sites (ie. sequence variability) whereas real time PCR allows to quantify the initial DNA used as a template for amplification.
It sounds like you are very new to molecular biology. I suggest you start by doing some background reading on different techniques and their applications.
RFLP is simply a description of regions of DNA that do (or don't) have restriction sites for restriction endonuclease enzymes. It has lots of potential applications (forensics, DNA fingerprinting, crop breeding, genotyping, etc.). It's pretty easy to learn, I even use it with my undergrads.
qPCR is a very sensitive way to measure DNA (or cDNA) copy numbers. Again, it has LOTS of potential applications (disease diagnosis, gene expression, etc.). It's HARD to master and you'll spend much longer trouble-shooting than actually doing your experiment.
Which one you choose depends on your research question.
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