Building a diverse CRISPR gRNA library using arrayed-synthesized oligo pools

CRISPR gRNA libraries provide a great tool for genome-wide and gene-focused applications, such as studying molecular targets or therapeutic target screening. However, to have a successful CRISPR gRNA library, the quality of the gRNAs is important and requires ultimate control over the synthesis process. This application note will demonstrate how using an arrayed, semiconductor-based oligonucleotide synthesis platform for synthesis of gRNAs creates a reliable, pooled CRISPR gRNA library for rapid high-throughput screening of molecular targets.

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In this application note, discover how using a pool of arrayed-synthesized gRNAs allows easy construction of a diverse CRISPR gRNA library with complete coverage and uniform distribution, maximizing your screening efficiency.

This application note highlights

Arrayed-synthesis of gRNAs creates a pool of gRNAs with 100% diversity

Precise synthesis of gRNAs creates a CRISPR gRNA library with complete coverage and uniform distribution of all desired sequences

Guidelines for improving gRNA design and selecting cloning vectors.

Advantages of using array-synthesized, double stranded DNA capture probes in NGS targeted sequencing

The key to excellent target enrichment relies on the use of high quality capture probes and creating optimized target enrichment probes improves the efficiency of targeted NGS and reduces the number of sequencing reads needed to obtain high-confidence data. In this application note, learn how array-synthesized, double stranded DNA capture probes can provide improved and consistent target capture in NGS targeted sequencing.

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This application note highlights

Double stranded DNA probes can provide deep, uniform sequence coverage & high targeting efficiency

Excellent target capture reproducibility using double stranded DNA probes

Comparable quality to single stranded DNA probes

Assembly of precise mutant libraries using arrayed-synthesized oligos for protein optimization

Mutant libraries are used to identify critical residues within a protein or to optimize protein function. Traditional methods for mutant library construction, such as using error-prone PCR or degenerate oligos, introduce mutations across the sequence space, however, have limited control over the codons introduced and variants can incur significant codon bias. The result is a large library with poor variant representation and a large screening burden to capture the entire variation of the library.

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In this application note, learn how using an arrayed-synthesized oligos can eliminate codon bias and poor variant representation to construct a well-designed, diverse mutant library.

This application note highlights

Array-synthesis of oligos provides precise control over codon usage in mutant variants

Elimination of codon bias and poor mutant variant representation

Comparison of mutant libraries constructed with NNK degenerate oligos & arrayed-synthesized oligos

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