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Library Construction

VectorBuilder has extensive experience in designing and constructing high-complexity pooled libraries for large-scale functional screens. Whether you're exploring gene function, conducting CRISPR or RNAi screens, investigating regulatory elements, or performing cell lineage analyses, our team of experts is dedicated to deliver high-quality libraries that are tailored to your specific research goals. Our pooled libraries are fully validated by next-generation sequencing (NGS) so you know exactly what you get.

Premade Pooled Libraries

Workflow for Library Construction

workflow for library construction
Library design

Designing an optimized library is critical to a successful screening experiment. Our technical team can help you design a library vector that is compatible with an efficient and error-less cloning process, contains proper marker or barcode sequence for versatile downstream screening experiments, and has high signal-to-noise ratio for robust results. Depending on your research needs, you can choose from our various validated delivery systems, such as lentivirus, AAV, piggyBac, non-viral regular plasmid, and adenovirus.

For CRISPR or RNAi screen, if you need assistance in designing gRNA or shRNA sequences for your list of target genes or genomic regions, VectorBuilder can help you design them using our established algorithm and computational pipeline. For 2-vector CRISPR libraries, we can also construct and package Cas9 expression vectors that are suitable for your screening strategy.

Pooled library cloning

To obtain DNA variants to be inserted in the vector backbone, we can use de novo oligo synthesis, directed evolution, or combinatorial approach. The variants are PCR amplified for a minimal number of cycles and in-parallel cloned into the desired vector backbone with high coverage. E. coli cells harboring library plasmids are propagated and harvested in exponential growth phase to make glycerol stocks. To assess the coverage and uniformity of the library, NGS analysis is performed on the plasmid pool before proceeding to virus packaging.

Virus packaging of pooled library

You may need to deliver the pooled library into cells via viral transduction, such as infecting with lentivirus or AAV. VectorBuilder has developed a range of proprietary technologies and reagents that have greatly improved virus packaging protocols for pooled libraries in terms of titer, purity, and uniformity. We can deliver the ready-to-use viral libraries for your screening experiments with a fast turnaround.

Click to view more information of virus packaging services  
NGS deconvolution

VectorBuilder can help you identify hits by NGS for your pooled library screens. You can simply send the genomic DNA of your samples to VectorBuilder, and we will prepare the NGS libraries, perform high-throughput sequencing, analyze the data, and deliver the accurate read counts in each sample to you in just a few weeks.

Service Details

Price and turnaround Price Match
Service Module Brief Description Price (USD) Turnaround
Library design Library vector backbone design for CRISPR, RNAi, barcode, enhancer/promoter, mutation, AAV capsid, and more pooled libraries, and gRNA or shRNA sequence design for your target genes/regions. Free 1-4 days
Pooled library cloning (including oligo synthesis and NGS validation) Massive parallel cloning of DNA variants into desired vector backbone, followed by preliminary QC by Sanger sequencing and full QC by NGS. The default deliverable is E. coli glycerol stock. From $2,300 5-8 weeks
Virus packaging of pooled library Please click here to view detailed info of virus packaging services. The price for packaging library is 1.5-fold of the price for packaging single vector.
NGS deconvolution of post-screening sample Includes NGS library preparation from genomic DNA of screened cells, Illumina sequencing (>500x coverage), and data analysis. From $320 per sample 3-5 weeks
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How to Order

Customer-supplied library plasmid pool

If the customer-supplied premade plasmid pools are used, please send us the materials following the Materials Submission Guidelines. Please strictly follow our guidelines to set up shipment to avoid any delay or damage of materials. All customer-supplied materials undergo mandatory QC by VectorBuilder which may incur a $100 surcharge for each item. Please note that production may not be initiated until customer-supplied materials pass QC. For customer-supplied premade plasmid pools, we cannot provide any guarantees regarding the complexity or uniformity of the library.

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