Oligo Pool—FAQs

Q1: How to connect the oligo pool to the corresponding vector after synthesis?

A1: 1. Design primers for PCR amplification of the oligo pool, with primers selected from the conserved region (15-20 nt). Additionally, when designing primers, consider the ligation method with the vector. If the ligation method is restriction enzyme digestion, add corresponding restriction sites and protective bases outside the conserved region. If the ligation method is homologous recombination, identify the homologous arms of the vector within the conserved region. Furthermore, ensure that the Tm values and lengths of the upstream and downstream primers do not differ significantly.

2.Enzymatic ligation method: Perform enzymatic digestion on PCR recovery products and vectors, followed by ligation with T4 ligase; Homologous recombination method: Use specialized enzymes for homologous recombination ligation, paying attention to the enzyme's operating temperature (PCR instrument may be utilized when necessary).

Q2: How to maintain sequence homogeneity to the greatest extent possible?

A2: During oligo pool synthesis, since the chip capacity is fixed, the number of primers should be doubled first when there are available positions. If the remaining space is insufficient for a full cycle, avoid random supplementation and maintain consistency. In PCR, primers bind randomly to the template, so minimize the number of cycles to prevent excessive amplification of certain bands due to repeated cycles, which can lead to redundancy. Reducing the number of cycles means fewer PCR products will be generated. Therefore, to ensure sufficient library capacity, the PCR system should be expanded accordingly.

Q3: The Correlation and Core Advantages of Oligo Pool with Trimer Codon Technology

A3: The core advantage of Trimer Codon technology lies in its use of trimeric nucleotide blocks (Trimer) instead of single nucleotides for synthesis, enabling precise, efficient, and diverse programming at the amino acid level in gene library construction. This is analogous to using "integrated bricks" (codons) rather than "scattered sand" (single nucleotides) in construction, fundamentally achieving direct editing of protein sequences. In summary, Trimer Codon elevates the unit of synthetic operations from "nucleotides" to "codons," significantly surpassing traditional methods in terms of precision, controllability, and efficiency. This makes it particularly suitable for directed protein evolution, antibody engineering, and enzyme optimization. The synthesis of oligo pools employs Trimer Codon technology, with its distinctive application being the construction of mutant libraries.

Q4: Camde Biotech's research on oligo pools

A4: In the application of oligo pools, Camde Biotech currently specializes in constructing customized peptide libraries, scFv antibodies, VHH antibody mutant libraries, and can also be used for creating customized aptamer libraries.