With the support of our advanced protein engineering platform established for many years, Creative BioMart provides a variety of methods to select the best mutation sequence or sequence library to identify mutants that contain the desired characteristics defined by protein engineering goals. Our scientists provide you with fully customized gene amplification service based on the plymerase chain reaction technology.

Polymerase chain reaction (PCR) is a technique used to exponentially amplify specific target DNA sequences, allowing individual sequences to be separated, sequenced, or cloned among many sequences. This technique involves the enzymatic amplification of nucleic acid sequences through repeated cycles of denaturation, oligonucleotide annealing, and DNA polymerase extension. PCR has completely changed the practice of DNA technology because it can easily generate almost any nucleic acid sequence in vitro with relatively high abundance. PCR can also change the sequence of individual DNA fragments. In many cases, this method has advantages over traditional DNA cloning and analysis methods because it is faster, simpler, and more flexible. At present, this technology has many applications in basic and applied research of medicine, agriculture, environment and bio-industry.

Services

In protein engineering, standard molecular biology techniques are used to manipulate DNA sequences that encode proteins, in which specific base pairs of protein-coding genes are identified and modified strategies using overlapping PCR. Our polymerase chain reaction gene amplification service allows rapid amplification of the best gene mutation sequence to generate mutants with the required characteristics. It can also facilitate the cloning of large or complex DNA fragments into related cloning vectors without the use of internal restriction endonuclease sites.

Fig 1. Overview of a Polymerase Chain Reaction Cycle. (Image Source: BioNinja)

Methods of Polymerase Chain Reaction

• Reverse transcription polymerase chain reaction: the thermostable DNA polymerase used for basic PCR requires a DNA template, so this technique is limited to the analysis of DNA samples.
• Hot-start PCR: hot-start PCR is a commonly used technique to reduce non-specific amplification due to assembly amplification reactions at room temperature.
• Quantitative real-time PCR: the use of fluorescently labeled oligonucleotide probes or primers or fluorescent DNA binding dyes to detect and quantify PCR products allows real-time quantitative PCR.
• Quantitative end-point PCR: the amplification efficiency is reduced due to inhibitors, competitive reactions, and depletion of substrates. As the number of cycles increases, the amplification efficiency decreases, and finally a platform effect appears. Quantitative PCR requires measurement before the plateau period so that the relationship between cycle number and molecule is relatively linear.

Applications of Polymerase Chain Reaction

The polymerase chain reaction gene amplification service is not limited to selective isolation and DNA amplification in protein engineering. It also includes the diagnosis and monitoring of genetic diseases, genetic fingerprint analysis of DNA analysis, and nucleic acid testing of detected pathogens for the diagnosis of infectious diseases.

Service Principle

Creative BioMart is based on innovative, pragmatic and honest, adhering to the tenet of "Quality is our life, providing customers with the best quality service", providing customized services to customers around the world.

We will be glad to discuss details of intended interaction studies with you and develop experimental strategies/methods tailored to your requirement. Our customer service representatives are enthusiastic and trustworthy 24 hours a day, Monday to Friday. Please do not hesitate to contact us for more information or to discuss in detail, if you are interested in our services.

References

1. Yamabhai M. (2009) Sticky PCR: A PCR-based protocol for targeted protein engineering. Biotechnol J. 4(4): 544-553.
2. Rashtchian A. (1995) Novel methods for cloning and engineering genes using the polymerase chain reaction. Curr Opin Biotechnol. 6(1):30-36.

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