Wednesday 10 April 2024

Multiplex PCR

A method called multiplex PCR, or polymerase chain reaction, enables the simultaneous amplification of many DNA fragments in a single operation. Multiple target sequences can be found and identified in a single reaction tube, which can be beneficial in terms of money, time, and sample preservation.



Here's a brief overview of how it works:

  1. Primer Design: Primers are short DNA sequences that bind to specific regions flanking the target sequences. In multiplex PCR, multiple sets of primers are designed, each specific to a different target sequence.

  2. Template DNA: The DNA sample containing the target sequences is added to the PCR reaction mix.

  3. PCR Reaction: The PCR reaction mix contains DNA polymerase, nucleotides, buffer, and the designed primers. The reaction goes through cycles of denaturation, annealing, and extension.

    • Denaturation: The double-stranded DNA template is heated to separate the two strands, resulting in single-stranded DNA.

    • Annealing: The reaction is cooled to allow the primers to bind (anneal) to their complementary sequences on the target DNA.

    • Extension: The temperature is raised, and the DNA polymerase extends the primers, synthesizing new DNA strands complementary to the target sequences.

  4. Detection: The amplified DNA fragments are detected and analyzed. Various methods can be used for detection, including gel electrophoresis, fluorescence detection, or real-time PCR.

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