PCR and RFLP are two different techniques that can be used to amplify or identify the differences between organisms. To understand this better, let's see what each of these techniques does.
In PCR, primers are designed to recognize the target sequence. In RFLP, restriction enzymes that recognize specific sequences in DNA fragments are used.
In PCR, a primer is an oligonucleotide that binds to a specific region of DNA and initiates DNA synthesis by adding deoxyribonucleotides onto the 3' end of an oligonucleotide chain. This process is repeated until there are enough copies of the target sequence so it can be detected using gel electrophoresis or polymerase chain reaction (PCR) detection methods such as fluorometry or ethidium bromide staining.
PCR products can be visualized by electrophoresis on agarose gels or on an appropriate matrix for capillary electrophoresis. RFLP products are visualized by electrophoresis on a gel. PCR products can also be visualized by capillary electrophoresis.
RFLP is a technique used to detect polymorphisms within cells. Polymorphism is the variation of DNA among individuals due to mutations that make it possible to identify individuals from their genetic material .
It relies on restriction enzymes, which are proteins that recognize and cut specific sequences in DNA (see figure below).
Restriction enzymes are not specific for one site in a particular chromosome, but rather have different sites they recognize on different chromosomes. When a particular restriction enzyme cuts a chromosome at its specific recognition site, it will produce two fragments with blunt ends called sticky ends (shown above). These sticky ends allow an RFLP reaction involving these fragments to occur more easily.
In contrast, PCR is a method that amplifies a region of DNA for many different purposes such as diagnosis and forensic analysis. PCR can be used to identify victims of crimes or natural disasters by comparing their DNA to that of their family members. In addition to this, you can also use PCR to diagnose diseases such as cancer and Huntington’s disease by amplifying specific regions of the genome that are associated with these diseases.
The main difference between these techniques is in the type of DNA that is analyzed to obtain information about its sequences. PCR, for example, can be used to amplify any DNA sequence (5’-GATGTACAGTCAATATCACGGCTTTTTCCTAACCATGCTCCCAGCCCAATTCT-3’). In contrast, RFLP depends on detecting polymorphisms at restriction enzyme sites.
PCR (polymerase chain reaction) is a technique used to amplify DNA. It is used in many applications, including DNA sequencing and genetic fingerprinting
It is also commonly used as a tool to detect the presence of specific DNA sequences in a sample.
RFLP uses restriction enzymes to digest the DNA of interest and the resulting fragments are separated by size on agarose gel. Whereas PCR is a method that helps to amplify a specific piece of DNA for analysis, RFLP does not amplify the sequence.
In PCR, the DNA primer is annealed complementary to the target gene's template.
The DNA polymerase synthesizes the primer in vitro by copying a segment of dsDNA that is complementary to a segment of ssDNA. The resulting double-stranded product is called the "hairpin."
Directional Cloning
In directional cloning, the DNA is amplified in one direction only. The DNA sequence of interest should be present at both ends of the fragment. A small amount of DNA is isolated and cut with restriction enzyme(s). The resulting fragments are placed in a vector that has been treated with the same restriction enzymes to allow for ligation. Directional cloning works well for amplifying specific regions of the genome because it does not produce any unwanted products due to replication from both strands (unwanted products can occur when using non-directional methods).
The main difference between PCR and RFLP is that PCR requires more technical skills and expensive equipment to perform the experiment. RFLP only needs a few reagents and can be performed on a wide range of samples, while PCR requires expensive equipment such as heat sealers and thermal cyclers.
Another difference between these two techniques is that PCR is more sensitive than RFLP, because it can detect small amounts of DNA with high sensitivity; whereas in RFLP you need a lot of DNA for amplifying your sample before you can detect the mutations in your sample.
In addition, another advantage of using PCR instead of RFLP could be its specificity; this means that if we are testing for a particular mutation in our sample, then it will give us exactly what we want without any false positives or negatives coming up during our test run infront us!
The main difference between PCR and RFLP is that PCR amplifies the DNA wherease RFLP identifies the difference between organisms by using restriction enzymes.
RFLP requires less expensive equipment and technical skills than PCR.
These techniques have many uses in genetics and medicine. The main advantages of PCR are the ability to amplify any DNA fragment, regardless of its size or sequence. RFLP is limited by the restriction enzyme recognition sites present in the genome, but it can be used for haplotype analysis (detection).