CEDT Equipment ( PCR INSTRUMENT )
Instrument PCR- Mastercycler
Manufacture Eppendrof
Model Mastercycler pro- Vapo.protect

Purpose of Instrument
  • The polymerase chain reaction (PCR) is a biochemical technology in molecular biology to amplify a single or a few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence.
  • PCR is now a common and often indispensable technique used in medical and biological research labs for a variety of applications. These include DNA cloning for sequencing, DNA-based phylogeny, or functional analysis of genes; the diagnosis of hereditary diseases; the identification of genetic fingerprints; and the detection and diagnosis of infectious diseases.
  • The method relies on thermal cycling, consisting of cycles of repeated heating and cooling of the reaction for DNA melting and enzymatic replication of the DNA. Primers (short DNA fragments) containing sequences complementary to the target region along with a DNA polymerase are key components to enable selective and repeated amplification.
  • Most PCR methods typically amplify DNA fragments of between 0.1 and 10 kilo base pairs (kb), although some techniques allow for amplification of fragments up to 40 kb in size.
  • PCR can be extensively modified to perform a wide array of genetic manipulations.
  • Typically, PCR consists of a series of 20-40 repeated temperature changes (called cycles) with each cycle commonly consisting of 3-4 discrete temperature steps.
  • Typically, PCR consists of a series of 20-40 repeated temperature changes (called cycles) with each cycle commonly consisting of 3-4 discrete temperature steps. These steps are :-
    1. Initialization step: This step consists of heating the reaction to a temperature of 94–96 °C.
    2. Denaturation step: This step is the first regular cycling event and consists of heating the reaction to 94–98 °C for 20–30 seconds.
    3. Annealing step: The reaction temperature is lowered to 50–65 °C for 20–40 seconds.
    4. Extension/elongation step: Depending on DNA polymerse, the temperature of this steps will usulally be between 70–80 °C.
Last Update
11/27/2014 10:24:30 PM