What are the different methods of PCR?

What are the different methods of PCR?

Some of the common types of PCR are;

  • Real-Time PCR (quantitative PCR or qPCR)
  • Reverse-Transcriptase (RT-PCR)
  • Multiplex PCR.
  • Nested PCR.
  • High Fidelity PCR.
  • Fast PCR.
  • Hot Start PCR.
  • GC-Rich PCR.

Which PCR method has the highest specificity?

It is concluded that double-step PCR employing nested primers is superior to other mycoplasma detection methods in many respects: simplicity and speed, high specificity and extreme sensitivity, objectivity and accuracy.

What is the most specific PCR?

Many of these measures critically depend on the timely and accurate diagnosis of virus-infected individuals. Real-time reverse transcription polymerase chain reaction (RT-PCR) is the most sensitive and specific assay and therefore preferred [2,3].

What are the 4 types of PCR?

Types of PCR

  • Real-time PCR.
  • Quantitative real time PCR (Q-RT PCR)
  • Reverse Transcriptase PCR (RT-PCR)
  • Multiplex PCR.
  • Nested PCR.
  • Long-range PCR.
  • Single-cell PCR.
  • Fast-cycling PCR.

What is the principle of nested PCR?

Nested PCR is a modification of PCR designed to increase the sensitivity and specificity of the assay reaction. It involves the use of two primer sets directed against the same target and two successive PCR reactions.

What is the difference between real time PCR and PCR?

What is PCR and how is it different from real time RT–PCR? RT–PCR is a variation of PCR, or polymerase chain reaction. The two techniques use the same process except that RT–PCR has an added step of reverse transcription of RNA to DNA, or RT, to allow for amplification.

How does a nested PCR work?

Nested polymerase chain reaction involves two sets of primers, used in two successive runs of polymerase chain reaction, the second set intended to amplify a secondary target within the first run product. This allows amplification for a low number of runs in the first round, limiting non-specific products.

Is nested PCR quantitative?

Nested qPCR, in which the product of the initial conventional PCR is used as the template for a second amplification with quantitative real-time PCR, has been used successfully to increase assay sensitivity in the diagnosis of tuberculous meningitis [16], respiratory viruses [17] and, more recently, Plasmodium …

What are the differences between PCR RT-PCR and RT qPCR?

QPCR and RT-PCR are both terms used in biotechnology and utilized for the production of multiple copies of DNA. RT-PCR is used to amplify the reversed transcription of the DNA code; QPCR measures the amplification. 3. RT-PCR is for amplification, while qPCR is for quantification.

Why nested PCR is preferred?

Nested PCR is well suited when only very small amounts of the target DNA are present in comparison to the total sample amount of DNA. Then two PCR are performed consecutively. The formula provides PCR the target gene is amplified beside unwanted sequence regions as a result of nonspecific binding of the primer.

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