Gene synthesis is a vital technique in molecular biology, allowing scientists to artificially create double-stranded DNA. This process typically involves assembling small segments of DNA or amplifying known samples of genetic material using the raw nucleobases guanine, cytosine, adenine, thymine, and uracil as starting materials. However, synthesizing high GC-content sequences, which refer to the proportion of guanine and cytosine in DNA and RNA molecules, can be challenging due to issues with secondary structure, mispriming, or mis-annealing.
GC-rich regions are known to facilitate base stacking, making them more stable than sequences with low GC-content. Additionally, secondary structures formed by high GC-content regions tend to be stable and more resilient to denaturation. However, sequences containing many guanine repeats can lead to complicated inter-strand folding due to hydrogen bonds between adjacent guanines, further complicating gene synthesis.
Temperature optimization is also closely related to gene synthesis, with primer GC ratio strongly influencing the predicted annealing temperature of DNA templates. High GC-content typically requires a high melting temperature, which can lead to mispriming or mis-annealing between the template and its complementary strand, leading to undesired and inaccurate gene products. This means that codon optimization – using synonymous codons to lower the GC-content and melting temperature – is a crucial aspect of gene synthesis.
Despite these challenges, Synbio Technologies is an expert in accurately synthesizing error-free DNA constructs to meet customers’ requests. They are capable of assembling multi-kilobase plasmids or entire genomes, even those with complex sequences such as hairpin structures, high GC ratio, high AT percentage, and multiple consecutive nucleotide sequences. Their success in synthesizing GC-rich constructs is critical for fully understanding and studying complex genes and even their non-coding segments with high conservation, with profound significance for biomedical research.
In conclusion, gene synthesis using high GC-content sequences can pose challenges in molecular biology, but Synbio Technologies has the expertise and tools to meet these challenges and produce high-quality, accurate DNA constructs. Their approach can help accelerate advancements in biomedical research and unlock new insights into the complex world of genetics.