Site Saturation Variant Library

January 10, 2024/in Partners, Synbio Libraries, Twist/by Harshita Sharma

PRODUCTS

Site Saturation Variant Libraries

Generate 99% of Desired Variants

The utilization of single-site variant libraries is proving invaluable for researchers aiming to delve into a protein’s sequence space and understand the intricate relationship between sequence, protein structure, and function. This innovative approach allows for a comprehensive exploration of variants, providing crucial insights into the molecular dynamics of proteins.

The construction of Twist Site Saturation Variant Libraries takes protein engineering to the next level. Leveraging advanced massively parallel oligonucleotide synthesis through Twist’s proprietary silicon-based DNA synthesis platform, these libraries ensure a precise and controlled crafting of variants. With complete mastery over codon usage, high uniformity at each site, and rigorous quality control verified through next-generation sequencing (NGS), these libraries offer researchers the flexibility to conduct experiments with ease. Whether opting for one position per well in a 96-well plate or pooling all positions in a single tube, this methodology enables the screening of 1 to 20 different amino acids at each position, further enhancing the adaptability and efficiency of protein engineering studies.

Site Saturation Libraries, particularly those constructed by Twist, revolutionize the exploration of sequence space in protein engineering by eliminating codon bias and preventing unwanted substitutions. In contrast to conventional methods like degenerate and NNK approaches, Twist Site Saturation Variant Libraries (SSVLs) offer a superior solution. Traditional methods, such as error-prone PCR and degenerate approaches, often suffer from poor repetitive yield, resulting in less than 50% full-length product in typical libraries. In comparison, Twist libraries yield more usable variants, significantly increasing the effective library size. The figure depicting the observed distribution of amino acids across 65 positions in a protein illustrates the efficiency of Twist SSVLs in maintaining the expected ratios of variants, showcasing their ability to provide highly uniform variant libraries.

Comparing key features, Twist SSVLs outshine error-prone PCR and degenerate approaches on various fronts. They eliminate sequence bias, utilize all 64 codons, prevent undesirable motifs, allow codon optimization, and avoid stop codons. This comprehensive set of advantages positions Twist SSVLs as a powerful tool for efficient sampling of a protein’s sequence space in screening assays, demonstrating their superiority in precision, reliability, and overall effectiveness in protein engineering studies.

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Synbio Libraries Portfolio

Site Saturation Variant Library

Combinatorial Variant Library

Spread Out Low Diversity Library

CAR Library

TCR Library

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Oligo Pools

January 10, 2024/in Partners, Synbio Oligo Pools, Twist/by Harshita Sharma

PRODUCTS

Oligo Pools

Twist Oligo Pools represent collections of single-stranded oligonucleotides synthesized using state-of-the-art silicon-based DNA writing technology. The innovative synthesis platform employed by Twist Bioscience allows for the simultaneous production of large quantities of high-quality and accurate oligos in each run. This capability facilitates the creation of complex and diverse oligo pools, specifically advantageous for applications such as CRISPR screening.

The process involves users submitting their oligo sequences to Twist Bioscience, which then synthesizes user-designed oligo pool libraries. This service enables researchers to dedicate their time to experimental work and discovery, with the synthesis handled by Twist.

The quality of Twist oligos is underscored by highly uniform synthesis, as evidenced by Next-Generation Sequencing (NGS) quality control data. A representative oligo pool containing 23,000 90mer oligos demonstrates remarkable uniformity, with the corresponding table indicating the metrics for this pool. Twist oligos are synthesized with minimal bias, ensuring high uniformity and complete representation of oligos in the pool.

Furthermore, sequencing analysis of Oligo Pools generated by Twist Bioscience is compared with those from an array-based competitor. The results highlight the superiority of Twist Oligo Pools, as they consistently contain 100% of the expected sequences and exhibit a higher percentage of correct sequences when compared to the competitor’s pool. This emphasis on accuracy and completeness in sequence representation makes Twist Oligo Pools a robust and reliable choice for various molecular biology applications.

New Cloned Oligo Pools

New Cloned Oligo Pools from Twist Bioscience represent a cutting-edge solution aimed at ensuring the foundational quality of oligo pools for successful experiments. Recognizing that errors during synthesis or cloning can adversely impact oligo pool quality by causing over and underrepresentation of desired sequences, Twist now provides an optimized cloning service. This service streamlines the experimental process by alleviating the need for researchers to navigate challenges such as testing PCR amplification conditions, selecting appropriate polymerases and primer pairs, and designing a cloning workflow.

The process involves just two steps. Users submit their oligo sequences to Twist Bioscience, and the company takes care of synthesizing, amplifying, and cloning the user-designed oligo pool libraries. This approach allows researchers to dedicate their time to experimentation and discovery while outsourcing the intricacies of oligo pool synthesis and cloning to Twist Bioscience.

Twist Bioscience emphasizes the high quality of its cloned oligo pools across all lengths, showcasing performance data for pools under 150 nucleotides and those up to 300 nucleotides in length, even with high GC content. The data highlight the uniformity achieved in both cases.

Twist’s cloned oligo pools not only exhibit low error rates and high uniformity but also boast low chimera rates. Chimeras, unwanted hybrid molecules resulting from suboptimal PCR conditions, can lead to improper amplification and recombination of different oligos within the pool. Twist Bioscience provides an illustration comparing an on-target cloned pool without chimeras to an off-target one containing chimeras, emphasizing the importance of avoiding undesired sequences.

The company asserts that its cloned oligo pools, free from chimeras, guarantee the highest quality and accuracy in experimental data. Comparative data between traditional amplification and Twist’s in-house amplification method further supports the efficacy of Twist’s approach in ensuring the integrity of oligo pools for diverse molecular biology applications.

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Other Panels

January 3, 2024/in NGS, Partners, Twist/by Harshita Sharma

PRODUCTS

Other Panels

Mitochondrial Panel

The mitochondrial genome encodes factors critical to energy production, which directly affects the energy state of the cell, tissue, and human individual. Pathogenic mutations of Mitochondrial DNA (mtDNA) are often implicated in a group of complex human diseases which can be difficult to diagnose such as metabolic & neurologic disorders as well as cancer. Interrogation of the human mitochondrial genome by targeted Next Generation Sequencing (NGS) can help investigators shed light on the genetic mechanism behind mitochondrial disorders.

The Twist Mitochondrial Panel is a fixed content panel designed to cover all 16,659 base pairs (bp) and 37 genes of the mitochondrial genome. This panel can be used as a standalone panel or as a spike-in with Twist’s Human Core Exome, Comprehensive Exome as well as Twist’s Custom Panels. Providing industry-leading coverage, uniformity, and flexibility, the Twist Mitochondrial Panel can help investigators enrich for, sequence, and analyze mtDNA variants contributing to disease including incidences of heteroplasmy.

Twist Alliance Diversity SNP Panel

Historical genotyping tools have long grappled with biases toward a limited number of ethnicities, resulting in underrepresentation of significant portions of the global population. Traditional platforms, like microarrays, exacerbate this issue by offering fixed content updated only periodically. To address these inefficiencies and promote equity, researchers can turn to target capture-based Next-Generation Sequencing (NGS) workflows with probe panels designed for diversity and flexibility.

Twist’s expanding Targeted Genomic Block Sequencing (GBS) portfolio includes the Twist Diversity SNP Panel, offering a flexible, ethnicity-neutral gold standard for genotyping. This panel can be used independently or as a spike-in with the Twist Human Comprehensive Exome panel. Its customizable design allows for additional probes to cover specific regions of interest, all synthesized on Twist’s DNA synthesis platform.

Developed in collaboration with the Regeneron Genetics Center, the Twist Diversity SNP Panel seamlessly integrates into existing fully automated exome processing workflows, providing base calls and imputed variants. Genotype imputations can be calculated using open-source tools, with an imputation guide available from Twist Customer Support for those starting with imputation analysis tools.

Note: While the Diversity SNP panel content has not been fully validated through the complete Twist workflow, users are encouraged to contact Twist Customer Support for recommendations on assay set-up.

Twist Alliance Pan-cancer Methylation Panel - 1.5 MB

Methylation sequencing plays a crucial role in detecting and tracing the origins of cancer cells, offering sensitivity in unraveling complex methylation patterns. The Twist Alliance Pan-cancer Methylation Panel – 1.5 MB, a collaborative effort between Twist and AnchorDx, is tailored to focus on cancer-specific methylation patterns across 31 different cancers. This custom target enrichment panel provides deep coverage of clinically relevant targets, enabling the study of methylation patterns crucial for early cancer detection and diagnosis, especially from tumor and liquid biopsy samples.

Constructed with Twist’s state-of-the-art oligonucleotide synthesis platform, the panel boasts high on-target rates and uniformity. It has undergone validation with the Twist Methylation workflow, utilizing liquid biopsy samples from breast cancer, colorectal cancer, and non-small cell lung cancer. The panel design, based on the TCGA database, covers 31 cancer types and 47 disease entities, featuring 13,090 probes targeting approximately 126k CpG sites. Notably, around 74% of the 12k differentially methylated regions (DMRs) overlap with CpG islands, showcasing the panel’s comprehensive coverage.

It’s essential to note that the Twist Alliance Pan-cancer Methylation Panel – 1.5 MB does not carry ISO-13485 certification.

Twist Alliance CNTG Exome - 41 MB

The Twist Alliance CNTG Exome – 41 MB, developed through collaboration between CENTOGENE and Twist, serves as a powerful tool for addressing the challenges associated with the diagnosis of rare diseases. Rare diseases often pose difficulties for physicians, leading patients on prolonged diagnostic journeys. DNA sequencing panels, such as this collaboration, offer comprehensive solutions, providing answers and resolutions for patients undergoing these diagnostic odysseys.

As a leader in rare disease diagnostics, CENTOGENE leverages its expertise in conjunction with Twist’s advanced omics technology to design target enrichment capture panels. The Twist Alliance CNTG Exome – 41 MB ensures highly uniform coverage of the entire exome, including full coverage of the mitochondrial genome. This panel significantly enhances diagnostic yield compared to standard whole exome sequencing, addressing a broad spectrum of disorders encompassing over 7,000 rare diseases.

The panel comprises a total of 483,000 probes, with components such as the Twist Human Comprehensive Exome (396.1K probes), Twist Mitochondrial Panel (139 probes), Twist Alliance Centogene Rare Disease Panel (78.8K probes), and Centogene Spike-In (7.8K probes). Notably, the Twist Alliance CNTG Exome – 41 MB does not carry ISO-13485 certification, emphasizing its designation for research use only and not for diagnostic procedures.

Twist Alliance CNTG Rare Disease Panel - 7.6 MB

The Twist Alliance CNTG Rare Disease Panel, designated for research use only and not for diagnostic procedures, focuses on providing a more streamlined solution for researchers targeting rare diseases. With over 2,500 relevant genes associated with rare diseases, this panel ensures comprehensive coverage, encompassing ≥99.5% of targeted regions within the CNTG Exome footprint.

Containing 78.8K probes, the Twist Alliance CNTG Rare Disease Panel is designed to detect genes linked to various conditions, including rare treatable diseases, early onset childhood disorders, cardiac disorders, neurodegenerative diseases, skeletal abnormalities, and metabolic disorders. This lightweight tool offers researchers the flexibility to capture crucial rare disease coverage without the full exome footprint.

It’s important to note that the Twist Alliance CNTG Rare Disease Panel-7.6MB does not carry ISO-13485 certification, emphasizing its specific designation for research purposes.

Twist Alliance CNTG Hereditary Oncology Panel - 0.2 MB

The Twist Alliance CNTG Hereditary Oncology Panel, expressly designed for research use only and not intended for diagnostic procedures, focuses on providing a targeted solution for identifying genetic predisposition to hereditary tumors. This panel includes 72 carefully selected cancer-associated genes and covers ≥99.5% of targeted regions.

With applications ranging from identifying genetic predispositions in breast cancer to gastrointestinal tumors, Li-Fraumeni syndrome, MEN1, MEN2, paragangliomas, and more, the Twist Alliance CNTG Hereditary Oncology Panel offers a specialized tool for researchers delving into the genetic underpinnings of hereditary cancers.

It’s essential to note that the Twist Alliance CNTG Hereditary Oncology Panel- 0.2MB does not carry ISO-13485 certification and is exclusively designated for research purposes.

Twist Human Ancient DNA Panel

The Ancient Human DNA Panel, a collaborative effort with David Reich’s lab at Harvard University, is designed for studying genetic variation in ancient DNA specimens, typically obtained from bones and teeth. Targeting 1.35 million SNPs, this panel includes a subset covering all 1.23 million polymorphic sites in the human genome published in Fu et al. Nature 2015. This ensures compatibility with published data on numerous ancient individuals, fostering broad comparability.

Developed using Twist’s double-stranded capture probes and hybridization capture reagents, the panel demonstrates high genome coverage and reduced allelic bias. This optimization, as highlighted in Rohland, Mallick et al. Genome Research 2022, enables genome-scale data at significantly reduced sequencing costs compared to shotgun sequencing, thanks to the highly efficient enrichment of human DNA fragments.

It’s important to note that the Twist Alliance Ancient Human DNA Panel is not ISO-13485 certified, emphasizing its exclusive use for research purposes.

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NGS Portfolio

Custom Panels

Viral Panels

Methylation Detection