The PCR is one of the best techniques for marker assistant selection. Immuno PCR is a combination of real-time PCR and ELISA methods. The unique DNA sequence of a particular virus is targeted for the identification. Annealing- in The primer binds or anneals to its exact complementary sequence on a DNA during the annealing step. There is a range of different probe technologies available, all using fluorophores. Multiplex PCR. PCR or Polymerase Chain Reaction is a technique used in molecular biology to create several copies of a certain DNA segment. With the help of the Taq DNA polymerase, the dATP, dGTP, dCTP and dTTP binds at its complementary nucleotides on the growing DNA strand. Two sets of primers are used in two successive reactions. Nested PCR. The first set of primers allows a first amplification. It helps in analyzing the gene expression. The chance of cross-contamination is always high in the case of the PCR. well, it can not work at a higher temperature. The polymerase chain reaction is a highly sensitive biological technique. At 94ºC temperature, the double-stranded DNA opens up by breaking hydrogen bonds. Now put the tubes in the PCR machine one by one in the pre-set PCR protocol. In situ-PCR is yet another excellent method for rapid amplification of a sample DNA. 1.3 Nested PCR This PCR increases the sensitivity due to small amounts of the target are detected by using two sets of primers, involving a double process of amplification [15, 16]. The produc t of this PCR is subjected to a second PCR using the second set of primers. Primers are typically diluted in molecular grade water to a stock concentration of 100 μM (100 pmoles /μL). Multiplex PCR is a widespread molecular biology technique for amplification of multiple targets in a single PCR experiment. All three steps are repeated for 25 to 40 cycles and in each cycle the DNA becomes double. Quantitative PCR is used to measure the specific amount of target DNA (or RNA) in a sample. Nested PCR is a modification of PCR that was designed to improve sensitivity and specificity. DNA polymerase is the key enzyme that is present behind the whole process. Other utilities: PCR tubes, stands, pipettes, tips. Taq Polymerase can tolerate very high temperatures. For more detail on PCR buffer ingredients read the articles: The PCR machine is known as a thermocycler. The template must be DNA only. Replication is a process of DNA synthesis, however, for us mimicking replication in a lab isn’t possible. we can’t visualize a few DNAs that is why we need to amplify DNA. 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 […] The in silico PCR is a computational tool used to estimate or predict the results of actual PCR reaction. It is an enzymatic method and carried out invitro. The PCR technique is based on the enzymatic replication of DNA. Though the method is similar, the optimization must be required for developing different multiplex protocols. The principle of the Nested PCR DNA is that the product of the first PCR is used for the second amplification. The DNA polymerase adds about 1000bp/minute under optimum conditions. Nested PCR used two sets of Primers. Nested PCR involves the use of two primer sets and two successive PCR reactions. PCR uses the enzyme DNA polymerase that directs the synthesis of DNA from deoxynucleotide substrates on a single-stranded DNA template. In each step, different reactions occur. Furthermore, we will also discuss some of the important types of PCR used to enhance PCR results. First pair -amplify a fragment similar to a standard PCR. This is used for the amplification of multiple targets in a single PCR experiment. 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