Variant Libraries

Variant libraries are constructed by incorporating specific or random mutations into given DNA sequences, and play important roles in directed evolution, functional genomics, protein engineering, and drug screening. GCATbio offers a variety of options, including site saturation variant libraries, site scanning variant libraries, and combinatorial variant libraries. Our cutting-edge mMPS high-throughput DNA synthesis technology enables us to efficiently generate all the oligos required for precision variant libraries. We provide custom codon optimization based on downstream expression systems, tailoring the codon usage for each variant position and eliminating unwanted codons or premature stop codons. All libraries are high-throughput sequencing verified, with the target coverage reaching 100%.

Key Benefits

Precision synthesis

Enables precise control of codon ratio, avoiding unnecessary mutations and facilitating subsequent screening and characterization

Superior uniformity

Uniform distribution of amino acids and high library diversity prevent specific amino acid deficiencies or over-expression, which avoids downstream screening biases

High library coverage

Libraries of various sizes can all guarantee 100% coverage

High yield

The yield per oligo reaches the pmol level, significantly increasing yield compared to other array-based chip synthesis (fmol-level). With no need of additional amplification or enzyme treatment, the libraries can be directly used for downstream experiments

Precision site saturation variant library

One or multiple amino acids in the same domain within the target protein sequence can be mutated into the remaining 19/20 amino acids, with custom codon frequency at each position;

Only one position can be mutated at a time (each variant contains only one amino acid mutation);

A sub-library consists of variants with mutations at a given amino acid position.

Precision scanning saturation variant library

Consecutive amino acids in a given domain or the entire target protein sequence can be mutated into the remaining 19/20 amino acids, with custom codon frequency at each position;

Only one position can be mutated at a time (each variant contains only one amino acid mutation);

A sub-library consists of variants with mutations at a given amino acid position.

Precision combinatorial variant library

Consecutive amino acids in one or more domains within the target protein sequence can be mutated into the remaining 19/20 amino acids, with custom codon frequency at each position;

Multiple positions can be mutated simultaneously (each variant contains multiple amino acid mutations);

A sub-library consists of variants with simultaneous mutations at multiple amino acid positions within the same domain.

Precision Scanning Saturation Variant Libraries Support Genotype-Phenotype Association Studies

GCATbio provided end-to-end tailored services to design and synthesize an NNK scanning saturation variant library for target gene regions (100-300 bp). Customer requirements include adjustable codon distribution and proportional distribution of degenerate bases. After library synthesis, high-throughput sequencing analysis confirmed the uniform distribution of introduced 32 codons, with results indicating no significant codon preference at each site and 100% library coverage. This facilitated efficient screening in the downstream applications of genotype-phenotype association studies, garnering high praise from the customer.

Ordering

1. Please download the GCATbio_Variant Libraries Quote Request Form and email us, or directly upload your requirements online. A dedicated project manager will respond to your email within 24 hours and provide professional advice for building your library.

2. The GCATbio email system employs robust confidentiality measures to safeguard project details, ensuring strict confidentiality, and all project intellectual property rights belonging to the customer.

Variant Libraries