Genes can be anywhere between a few hundred and a few thousand base pairs long, therefore producing lengthy sections of nucleotide sequence was initially difficult for Gene Synthesis. The maximum length of oligos that can currently be produced using solid-phase oligonucleotide synthesis technique is 200 nucleotides. Polymerase cycling assembly and synthesis by ligation are the two main strategies for getting around this problem. Instead of depending solely on the entire synthesis of both DNA strands, polymerase cycling assembly (PCA) makes use of single-stranded templates. The synthesis of short oligos that alternate between the gene's two strands and overlap by 20–30 base pairs occurs.
Even after error correction, the result frequently still has numerous faults in the sequence, despite the fact that PCA makes it easier to synthesize big genes and genomes. Since the fragment oligos must span the whole sequence of both the sense and antisense strands, gene synthesis via ligation is only feasible for shorter genes and has a lower error rate than PCA.
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