Polymerase Chain Reaction Basics
Start with four main components of a Polymerase Chain Reaction
Polymerase chain reaction (PCR) is a method widely used to rapidly make millions to billions of copies of a specific DNA sample, allowing scientists to take a very small sample of DNA and amplify it to a large enough amount to study in detail. There are four main components required for a reaction: DNA template, DNA primers, free nucleotides called dNTPs, and DNA polymerase (e.g. U-Taq DNA Polymerase). At SBS Genetech, we are at the forefront of offering comprehensive solutions for PCR-Related products based on our various platforms.
PrimeTaq™ HiFi PCR Polymerase, which is derived from high fidelity DNA polymerase and added with enhanced extension structure, has super high fidelity (~ 280 times of Taq DNA Polymerase), long fragment amplification ability, and higher yield. The polymerase can easily amplify 8 kb genomic DNA, 20 kb λDNA, and 8 kb cDNA. The polymerase has an extension rate of more than 6 kb/min.
The Development of PCR
From basic chemical reaction to advanced statistical analysis
The 1st Generation
Traditional PCR
By comparing the intensity of the amplified PCR product on a gel to standards of a known concentration (usually using DNA Markers) at the end of the PCR cycles, the results of qualitative detection are obtained. Traditional PCR suffers from poor precision and low sensitivity.
The 2nd Generation
Real-time PCR
By collecting data during the exponential growth phase of PCR (e.g. signal from the fluorescent reporter), the quantity of the PCR product can be measured as it is directly proportional to the amount of starting template nucleic acid. Real-time PCR is very precise and specific.
The 3rd Generation
Digital PCR
By measuring the fraction of negative replicates, the absolute copies (e.g. viral load) can be determined by Poisson statistical algorithm. Digital PCR provides a novel solution for absolute quantification and there are no references or standards required.
Advanced Solutions
With PrimeTaq™ series, everything becomes easy and "direct"
Direct PCR of DNA from Crude Sample
By PrimeTaq™ HotStart Direct PCR DNA Polymerase
PrimeTaq™ HotStart Direct PCR DNA Polymerase has hot start property and high tolerance to many PCR inhibitors. High tolerance delivers convenience for DNA amplification by allowing PCR directly from samples without prior DNA purification. With hot start property, the polymerase is 100% inactive below 50°C and can be completely recovered only after heating at 95°C for 5 min. Therefore, the system can effectively inhibit non-specific PCR amplification, greatly improving the specificity and sensitivity. The polymerase has 5 '- 3' exonuclease activity, without 3 '- 5' exonuclease activity.Direct PCR of RNA from Crude Sample
By PrimeTaq™ HS DP DNA/RNA Polymerase
PrimeTaq™ HS DP DNA/RNA Polymerase has hot start property and high tolerance to many PCR inhibitors. High tolerance delivers convenience for both DNA and RNA amplification by allowing PCR directly from samples without prior purification. With hot start property, the polymerase is 100% inactive below 50°C and can be completely recovered only after heating at 92°C for 5 min. Therefore, the system can effectively inhibit non-specific PCR amplification, greatly improving the specificity and sensitivity.Direct Quantification of DNA from Crude Sample
By PrimeTaq™ Probe qPCR MasterMix
PrimeTaq™ Probe qPCR MasterMix, which premixes reagents including chemically modified PrimeTaq™ HotStart Direct PCR DNA Polymerase, reaction buffer, and dNTPs. This MasterMix does not contain ROX dyes, which is compatible with a wide range of fluorescent probes and a variety of quantitative PCR instruments. In addition, this MasterMix has great performance in multiplex quantitative PCR (multiplex qPCR) assays, enabling a minimum of 4-plex quantitative PCR assays.
Direct Quantification of RNA from Crude Sample
By PrimeTaq™ Probe One-Step RT-qPCR MasterMix
PrimeTaq™ Probe One-Step RT-qPCR MasterMix contains PrimeTaq™ HS DP DNA/RNA Polymerase, which has hot start property and high tolerance to many PCR inhibitors. High tolerance delivers convenience for both DNA and RNA amplification by allowing PCR directly from samples without prior purification. With hot start property, the polymerase is 100% inactive below 50°C and can be completely recovered only after heating at 92°C for 5 min. Therefore, the system can effectively inhibit non-specific PCR amplification, greatly improving the specificity and sensitivity.
SBS Genetech is a long-term sponsor of
Cold Spring Harbor Laboratory
