Full-Spectrum Library Construction

Mutant Library Construction Service

Build the genetic diversity every downstream strain-improvement workflow depends on — from million-clone random pools to single-base targeted edits and genome-wide perturbation maps. Part of our Microbial Strain Engineering & Fermentation Optimization Platform.

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10⁶–10⁷

mutants / run

3 approaches

random · targeted · genome-wide

organism categories

Schematic of a mutant library construction framework spanning random, targeted, and genome-wide mutagenesis converging into a screening-ready library.

Full-Spectrum Coverage

random to genome-wide

Random, targeted & genome-wide — one integrated workflow

Goal-matched resolution from single-base edits to 10⁷ clones

Screening-ready for direct handoff

Overview

Three Approaches, One Library-Building Engine

Every better strain starts with the right kind of genetic diversity. We let the goal choose the method.

Industrial strain development begins with genetic diversity, but the optimal way to create it depends on what you are looking for. Creative BioMart Microbe delivers a full-spectrum Mutant Library Construction service that scales resolution to the question: broad random diversity for discovery, precise edits at known sites for optimization, and systematic genome-wide coverage for functional mapping.

Random mutagenesis applies unbiased physical, chemical, and polymerase-based mutagenesis to search the widest possible phenotype space. Targeted mutagenesis introduces changes at defined loci through site-directed, saturation, and combinatorial methods. Genome-wide methods deploy transposon insertion, recombineering, and CRISPR/Cas genome-wide libraries to interrogate entire genomes at population scale.

Constructed libraries are formatted for immediate processing by our High-Throughput Screening Service, enabling rapid identification of improved producers without reformatting or rework.

Our integrated pipeline spans library construction through single-cell resolution phenotyping, ensuring seamless diversity preservation from mutagenesis to hit isolation.

Services

A Six-Step Path From Inquiry To Library

Each project follows a six-step workflow from initial consultation to documented delivery, with the mutagenesis approach selected to match your objective.

Project Consultation

Approach selection & goal review

Strain & Material Prep

Culture, plasmid, or DNA receipt

Mutagenesis / Engineering

Selected method applied

Recovery & Expansion

Stabilize and amplify clones

Quantification & QC

Scale, coverage, mutation rate

Delivery

Stocks + QC report

Service Details

Each approach trades breadth for precision — many programs combine broad random discovery with targeted refinement. Pick the one that matches your starting point and goal.

Random Mutagenesis

We apply physical agents (UV, ion beam, atmospheric-pressure plasma), chemical mutagens (EMS, NTG, sodium nitrite), and error-prone PCR to introduce unbiased, genome-wide diversity. Atmospheric-pressure plasma delivers a broad mutation spectrum at ambient temperature with high viability retention. Best for: exploratory diversity where the target is unknown.

Targeted Mutagenesis

We engineer precise changes at known loci using site-directed, saturation, and combinatorial mutagenesis. These methods localize diversity to a gene or active site of interest, accelerating enzyme and pathway optimization with minimal off-target background. Best for: enzyme and pathway optimization at defined sites.

Genome-Wide Mutagenesis

We deploy transposon mutagenesis (Tn5, Himar1 mariner, mini-Tn7), λ Red recombineering, and CRISPR/Cas genome-wide libraries to interrogate entire genomes. These tools enable systematic loss-of-function and gain-of-function mapping at population scale. Best for: functional genomics, essentiality, and pathway discovery.

Technique Coverage Matrix

Approach	Technique	Target Substrate / Representative Hosts	Deliverable
Random	Physical mutagenesis — UV, ion beam, atmospheric-pressure plasma	Bacteria, yeast, fungi, algae, actinomycetes	Mixed mutant pool
Random	Chemical mutagenesis — EMS, NTG, sodium nitrite	Bacteria, yeast, fungi	Mixed mutant pool
Random	Error-prone PCR	Bacteria, yeast, fungi (plasmid-borne expression)	Variant gene library
Targeted	Site-directed & saturation mutagenesis	Cloned genes, plasmids	Defined variant set
Targeted	Combinatorial mutagenesis	Multi-site constructs	Combinatorial library
Genome-wide	Transposon mutagenesis — Tn5, Himar1 mariner, mini-Tn7	Bacteria	Saturated insertion library
Genome-wide	λ Red recombineering	Bacteria (E. coli and relatives)	Scarless / precise mutant set
Genome-wide	CRISPR/Cas genome-wide libraries	Bacteria, yeast	Knockout / perturbation library

Service Specifications & QC Standards

01 · Instrumentation & Capability

Physical, chemical, and PCR-based random mutagenesis platforms.
Targeted editing by site-directed, saturation, and combinatorial mutagenesis.
Transposon, recombineering, and CRISPR/Cas genome-wide toolkits.
Viability, mutation-rate, and coverage assessment by plating and NGS.
Compatibility with bacteria, yeast, fungi, actinomycetes, and algae.
Optional whole-genome or amplicon sequencing of selected clones.
Micro-droplet encapsulation and fluorescence-activated sorting compatibility for rapid single-cell phenotype screening.

02 · Typical Data Range

Random library scale / run	10⁶–10⁷ clones
Targeted variant count	10¹–10³ per site
Genome-wide coverage	Near-saturation (≥90% ORFs where applicable)
Species coverage	5+ categories
Process conditions	Strain-dependent; method-matched

03 · Deliverables

Mutant library	Glycerol stocks / plates / plasmid pool
QC report	Viability, mutation rate, coverage
Sequence data	Optional WGS of clones; NGS of pools
Documentation	Method parameters, construct lists

Sample Requirements

What To Send Us

To initiate a mutant library construction project, please provide the following information about your starting material and target.

Required	Optional	Not Accepted
Pure, axenic starting culture or validated plasmid / DNA Strain name and taxonomic identification (or sequence for targeted work) Biosafety classification (e.g., BSL-1) and known sensitivities Target approach and selection direction (where known)	Prior mutagenesis or screening history Desired library scale or variant count Preferred preservation format Structural model or target residue list for targeted work	Contaminated or mixed cultures Uncharacterized pathogens (BSL-2 or above without clearance) Strains under material transfer or export restrictions Non-viable or heavily lysed samples

Recommended Starting Material By Scale

10⁶ clones	1–2 glycerol vials or a fresh slant
10⁷ clones	2–3 glycerol vials plus a backup slant
Targeted / genome-wide	Plasmid, genomic DNA, or listed target locus

Storage & Shipping: Ship cultures on dry ice for glycerol stocks or as stabilized agar slants at ambient temperature with cushioning. Avoid temperature excursions and do not ship under conditions that cause complete lysis. For targeted and genome-wide projects, ship dry DNA or plasmids with a documented sequence file. Contact our team via the contact form before dispatch for biosafety clearance.

Advantages

Why Teams Build Libraries With Us

Full-Spectrum Breadth

Random, targeted, and genome-wide methods are delivered from one platform, so you can move from broad discovery to precise optimization without changing vendors.

Goal-Matched Resolution

Generate a 10⁷-clone search space for unknown targets, or confine diversity to a single active site when the objective is already defined.

Multi-Species Compatibility

The workflow supports bacteria, yeast, fungi, actinomycetes, and algae across multiple organism categories.

Seamless Screening Handoff

Libraries are formatted for immediate processing by our high-throughput screening service, eliminating reformatting delays.

Documented, Heritable Performance

Published data demonstrate stable, heritable improvements across multiple generations, confirming mutation stability for downstream screening and fermentation.

Flexible Delivery & QC

Libraries are delivered as glycerol stocks, agar plates, or plasmid pools with a QC report and optional whole-genome or amplicon sequencing.

Single-Cell Screening Ready

Constructed libraries are compatible with micro-droplet and fluorescence-activated sorting platforms, enabling phenotype screening at single-cell resolution without reformatting or rework.

Customer Reviews

Trusted by Research Teams Worldwide

"The random mutagenesis library exceeded our coverage expectations. NGS-based quality control confirmed genome-wide diversity, and the seamless handoff to high-throughput screening allowed us to identify improved producers within weeks."

Dr. J. M.

Metabolic Engineer

USA

"Their targeted saturation mutagenesis at the enzyme active site yielded variants showing a four-fold improvement in catalytic efficiency. The accompanying QC documentation and sequence validation were exceptionally thorough."

Dr. S. C.

Enzymologist

USA

"From the initial consultation through to final delivery, the team maintained clear communication and met every milestone. The formatted library arrived ready for immediate downstream screening without any reprocessing."

Dr. A. N.

Research Director

Germany

"The CRISPR-based genome-wide knockout library achieved near-saturation coverage of our target organism's ORFs. The inducible repression system and validated gRNA design rules gave us confidence in the functional mapping results."

Dr. P. B.

Synthetic Biologist

USA

"We combined random atmospheric-pressure plasma mutagenesis with targeted refinement of the top-performing clone. The platform's ability to move from broad discovery to precise optimization without changing service providers streamlined the entire program."

Dr. H. W.

Biotechnology Consultant

Germany

"Library delivery included comprehensive QC metrics covering viability, mutation rate, and coverage estimates. Having this level of documentation upfront made it straightforward to justify the next phase of screening to our funding committee."

Dr. Y. K.

Principal Investigator

USA

Applications

Where Mutant Libraries Create Value

Illustration representing industrial enzyme production.

Industrial Enzymes

Activity, thermostability, pH tolerance

Illustration representing organic acid fermentation.

Organic Acids

Titer, yield, substrate range

Illustration representing biofuel production.

Biofuels

Substrate tolerance, yield

Illustration representing pharmaceutical protein production.

Pharmaceutical Proteins

Secretion, folding

Probiotics

Digestive enzymes, stability

Illustration representing antibiotic and secondary metabolite production.

Antibiotics & Metabolites

Yield, precursor use

Case Study

Proven Across All Three Approaches

Random Mutagenesis: Plasma Treatment Raises β-Glucanase In Trichoderma reesei

A high β-glucanase activity mutant was isolated from Trichoderma reesei by atmospheric-pressure plasma mutagenesis followed by Congo red transparent-circle screening and enzyme activity rescreening. In 500 mL shake-flask culture, the mutant reached peak β-glucanase activity of 45.12 U/mL at 96 h, compared with 28.88 U/mL for the parental CICC 2626 strain, a 56.23% increase. Enzyme activity remained stable across seven consecutive passages with no significant difference (P > 0.05), confirming heritable improvement. Comparative transcriptomics and metabolomics further linked the enhanced phenotype to upregulated hydrolase genes and altered carbohydrate and amino acid metabolism. This example demonstrates that a single round of random mutagenesis can generate a genetically stable, high-performing strain suitable for direct downstream screening and fermentation optimization.

Monitoring of T. reesei CICC 2626 and mutant strain growth and β-glucanase activity in a 500 mL shake flask under uncontrolled pH conditions. — Figure 1. Monitoring of *T. reesei* CICC 2626 and the plasma-derived mutant strain growth and β-glucanase activity in a 500 mL shake flask under uncontrolled pH conditions. (Wang, *et al.* 2025)

Genome-Wide Mutagenesis: CRISPRi Library Maps Genes In Saccharomyces cerevisiae

An inducible, genome-wide CRISPR interference library was constructed for Saccharomyces cerevisiae using yeast-optimized spacer design rules. The library targets all open reading frames with six to twelve gRNAs per gene and employs anhydrotetracycline-inducible repression to control the timing and scale of perturbations. Validation experiments demonstrated accurate recovery of known haploinsufficient genes with a receiver operating characteristic area under the curve of 0.90, and successfully identified enzymatic and regulatory genes in adenine and arginine biosynthesis from a single round of selection. Systematic analysis further refined guide RNA design parameters, including optimal positioning within 150 base pairs upstream of transcription start sites and nucleosome occupancy thresholds. These findings confirm that inducible genome-wide CRISPRi libraries can quantitatively map gene function across entire genomes with high precision and low false discovery rates.

Figure 2. High-throughput identification of haploinsufficient and dosage-sensitive genes. (Momen-Roknabadi, *et al.* 2020)

Pair With High-Throughput Screening

Libraries constructed here are immediately compatible with our screening service for rapid hit identification from 10⁶–10⁷ diversity, with optional micro-droplet single-cell resolution.

Explore Screening →

FAQs

Common Questions

Q: Which mutagenesis approach should I choose for my project?

Choose random mutagenesis when the target phenotype is unknown and you need maximum discovery space, targeted mutagenesis when optimizing a known gene or active site, and genome-wide methods for functional mapping of entire genomes. Many programs combine approaches.

Q: Can you combine random and targeted approaches in one project?

Yes. A common workflow uses random mutagenesis to surface lead phenotypes, then targeted site-directed or saturation mutagenesis to refine the responsible locus. We design the handoff so the targeted step builds directly on the random hit.

Q: What organisms are compatible with transposon and CRISPR genome-wide methods?

Transposon mutagenesis (Tn5, Himar1 mariner, mini-Tn7) is applied to bacteria, while λ Red recombineering and CRISPR/Cas genome-wide libraries extend to bacteria and yeast. Host suitability is confirmed during consultation.

Q: How large a mutant library can you construct?

A standard random run delivers 10⁶–10⁷ mutant clones, scaled to your screening capacity. Targeted projects deliver 10¹–10³ variants per site, and genome-wide projects achieve near-saturation coverage of the relevant open reading frames.

Q: Do you provide sequencing to characterize the mutations?

Optional whole-genome sequencing of selected clones and amplicon or pooled-next-generation sequencing of libraries are offered to support mutation-spectrum assessment, coverage estimation, and follow-on engineering.

Q: Can the constructed library be paired with your screening service?

Yes. Libraries are formatted for direct handoff to our High-Throughput Screening Service, which supports both plate-based and micro-droplet fluorescence-activated sorting for rapid hit identification without reformatting.

Q: How are mutant libraries delivered and preserved?

Libraries are delivered as glycerol stocks, agar plates, or plasmid pools with a viability report, method parameters, and an optional mutation-spectrum assessment, preserved for downstream screening or evolution.

Ready To Build Your Mutant Library?

Tell us your starting material and target phenotype—our team will select the optimal approach and design the screening handoff.

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Explore The Strain Engineering Platform

Mutant Library Construction

Diversity generation

High-Throughput Screening

Million-scale discovery

Strain Directed Evolution

Iterative enhancement

Adaptive Laboratory Evolution

Industrial robustness

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References:

Wang, N., et al. (2025). Multi-omics analysis of Trichoderma reesei mutant with high glucanase activity. Scientific Reports, 15, 25184.
Momen-Roknabadi, A., et al. (2020). An inducible CRISPR interference library for genetic interrogation of Saccharomyces cerevisiae biology. Communications Biology, 3, 723.

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