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 ligase chain reaction technology.

Ligase chain reaction (LCR) is a probe amplification technique different from PCR. Under the action of DN A ligase, it quickly amplifies DNA fragments by connecting two adjacent oligonucleotide strands complementary to the template DNA. Each cycle causes the target nucleic acid molecule to double. The amplification efficiency of LCR is comparable to that of PCR. Because of the rapid, specific and sensitive characteristics of LCR, as well as the ability to detect single base mutations, it has been used in molecular diagnosis of tumor gene mutations, and combined with PCR to improve its sensitivity.

Services

The main purpose of protein engineering is to study the structure and function of recombinant proteins. DNA ligase is mainly used to connect DNA fragments in vitro, and protein recombination is possible through the connection of different DNA fragments. Our ligase chain reaction gene amplification service uses ligase ligation oligonucleotides as distinguishing sequence variants to become targets in subsequent cycles and undergo logarithmic amplification. In addition, it also allows these two complementary pairs to be blunt-ended to each other at low frequency and used as amplification templates even if the target sequence does not exist in the original sample.

Fig 1. Mechanism of DNA assembly via ligase cycling reaction (LCR). (de Kok S, et al., 2014)

Advantages of Ligase Chain Reaction

Compared with PCR-based gene amplification systems, LCR-based gene amplification systems have the following advantages:

• Since there is no newly synthesized DNA, erroneously incorporated nucleotides will not replicate in the product, allowing the amplification of sequences that differ from those found in the target nucleic acid.
• The LCR reaction is also more specific for 3' nucleotides, allowing higher discrimination against mismatches at a single selected site. LCR is very useful for determining the nucleotide at a specific site.
• The LCR cycle has only two short steps, which can shorten the amplification time.
• The LCR can amplify small nucleic acids (36 to 60 nucleotides), and does not require high-quality large nucleic acids.

Applications of Ligase Chain Reaction

The polymerase chain reaction gene amplification service is not limited to the detection of specific nucleic acid sequences in protein engineering. It is also used to detect single-base mutations.

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. Tanabe M, Ishino Y, et al.. (2015) From Structure-Function Analyses to Protein Engineering for Practical Applications of DNA Ligase. Archaea. 2015: 267570.
2. de Kok S, Stanton LH, et al.. (2014) Rapid and reliable DNA assembly via ligase cycling reaction. ACS Synth Biol. 3(2): 97-106.

• Natural Gene Amplification Service
• Polymerase Chain Reaction Amplification Service
• Transcription-Mediated Amplification Service