The synthesis of long genes (genes with extended sequences) involves several technical challenges, some of which include:

1. Increased Error Rates: As the length of the gene increases, the error rate during the synthesis process may rise. This includes potential insertions, deletions, or substitutions in the synthesized base sequence, making accuracy maintenance more challenging for longer genes.

2. GC Content Deviation: Synthesis of long genes may result in GC content deviation. Ensuring a balanced GC content throughout the entire gene sequence can be challenging, potentially leading to structural instability or other issues.

3. Decreased Synthesis Efficiency: Synthesizing long genes may lead to decreased synthesis efficiency. Some steps in chemical synthesis can become more complex in longer sequences, requiring more time and resources.

4. Complex Secondary Structures: For RNA genes in particular, longer sequences may form complex secondary structures, which could pose challenges during synthesis and subsequent expression. Maintaining the integrity of these structures may require more advanced synthesis and handling methods.

5. Optimization of Synthesis Strategy: Synthesizing long genes necessitates the optimization of synthesis strategies to ensure both efficiency and accuracy. This may involve staged synthesis, optimizing reaction conditions, and incorporating appropriate validation and correction steps.

6. Challenges in Cloning and Expression: Cloning and expressing long genes may also present challenges. Ensuring the correct insertion of the entire long gene sequence and its stable expression in host cells are crucial considerations.

In summary, the synthesis of long genes involves increased technical complexity, requiring a comprehensive approach to address challenges in synthesis, cloning, and expression.

GenCefe’s professional team has extensive experience in synthesizing difficult genes. View  our case   studies   of  synthesizing a  13.7kb gene: https://www.gencefebio.com/GeneSynthesis/93.html