PCR is a technique used in the lab to rapidly make millions to billions of copies of a complete or partial section
of a specific DNA sample. PCR is based on the ability of DNA polymerase to synthesize new strand of DNA
complementary to the offered template strand.
Double-stranded DNA can be denatured and unrolled into single-stranded DNA under the action of various enzymes. With
the participation of DNA polymerase, it is replicated into the same two molecular copies according to the principle
of complementary base pairing.
Procedure
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Denaturation of template DNA
Heat the reaction chamber to 94–98℃ to separate the double-stranded template DNA to two single-stranded DNA
molecules.
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Annealing (renaturation) between template DNA and primer
Lower the temperature to make the primer pair with the complementary sequence of the template DNA single strand.
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Extension of primers
Synthesize a new semi-retained replication strand complementary to the template DNA strand according to the
principle of base complementary pairing and semi-reserved replication.
Fig. 1 The basic flow chart for PCR.
Features
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Simple to understand and use, and be able to produce results rapidly.
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Highly sensitive with the potential to produce millions to billions of copies of a specific product for
sequencing, cloning, and analysis.
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Figure out the sequencing of unknown etiologies of many diseases.
Limitations
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Must know the precise sequence upstream of the target region on each of the two single-stranded templates.
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The smallest amount of contaminating DNA can be amplified, resulting in misleading or ambiguous results.
Applications
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Medical Application
For genetic testing where a DNA sample is analyzed for the presence of genetic disease mutations.
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Infectious Disease Application
Provide immediate solution for a specific infection and detect any resistance to antibiotics or other
therapeutic methods.
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Forensic Application
Run genetic fingerprinting and confirm the blood or genetic relationship between a concerned child and his
biological parents.
Related Products
Thermal Cycler is a laboratory apparatus that amplify segments of DNA using PCR. A thermal cycler is also commonly called a DNA Amplifier, PCR Machine, or Thermocycler. The cycle normally lowers or raises the block’s temperature in preprogrammed discrete steps. A thermal cycler is important for a laboratory dealing in molecular biology and gene cloning.
Digital PCR Systems (dPCR) is incredibly useful in applications such as mutation detection, copy number variation, rare sequence detection, gene expression analysis, miRNA analysis and next generation sequencing sample quantification. It provides a precise measure of DNA molecules in each drop. In some cases, Digital PCR Systems (dPCR) must be used in combination with a 96-deep well reaction module.
Real-time PCR System can quantify DNA copies and enable experiments in gene expression, genetic variation, genotyping, and specific detection of rare targets, bacteria, and viruses. It measures signals generated by fluorescent probes that are proportional to DNA amplification, allowing accurate quantification. It is capable of quantifying very small amounts of DNA with good dynamic range.
A PCR workstation, also called a PCR hood, is a work space, enclosed on three sides, that provides a space for doing amplification of DNA and/or RNA. PCR hoods are used in biology and genetic labs so that there isn’t any cross contamination between samples. PCR workstations have no circulation, which helps to prevent contamination, and UV lights for sterilization.
Electrophoresis equipment applies an electric charge to molecules, causing them to migrate towards their oppositely charged electrode. The technique is found in all research and clinical laboratories utilizing DNA and protein applications, and is divided into gel and capillary techniques. Choice of electrophoresis equipment depends on the molecule of study, appropriate method of separation and downstream application.
STEMart provides you with a variety of PCR equipment or consumables to meet your various R&D and application needs. If you have any questions or requirements for PCR, please feel free to contact us.