20µG SS DNA Synthesis CRISPR Homology Directed Repair Template Single Stranded DNA Synthesis

Single-stranded DNA (ssDNA or ssODN) is proven to be the best CRISPR homology directed repair (HDR) template for creating gene knock-in, with high editing efficiency and reduced off-target integration. CELLFREE now offers high quality, sequence verified ssDNA for maximizing the editing efficiency of your CRISPR experiments.

CELLFRE service provides up to 5000 nt of full sequence-verified fragments quickly and affordably. Our fragments are derived from clonally purified double-stranded DNA (dsDNA), producing the highest quality results possible.

ADVANTAGES OF ssDNA SYNTHESIS FOR CRISPR GENE KNOCK-INS

• Lower cellular toxicity compared to dsDNA after cellular delivery
• Low off-target integration for more reliable gene knock-ins
• High specificity knock-in templates for homology directed repair
• High efficiency donors for targeted insertions and gene replacements
• Yield flexibility – Choose 3, 10 or 20 µg of lyophilized fragments.

TURNAROUND TIME

DELIVERABLES

• Sequence verified
• 3 µg, 10 µg, or 20 µg lyophilized DNA
• Certificate of Analysis (COA) including gel image, sequence trace data with alignment, and sequence files

Application

1) CRISPR based gene insertion, replacement, or correction

Double-stranded DNA (dsDNA) was traditionally used as HDR donor DNA templates, however, recent studies demonstrated that single-stranded DNA (ssDNA or ssODN) is the best HDR templates for CRISPR based gene insertion, replacement, and correction. When compared to double-stranded DNA donors, ssDNAs demonstrated significantly improved editing efficiency and specificity, as well as reduced off-target integration, especially in editing primary cells, stem cells, and developing transgenic animal model.
2) In vitro transcription
In vitro transcription (IVT) method describes the transcription of RNA molecules via RNA phage polymerase (T7, T3 or SP6) using DNA templates in vitro. Three different types of DNA templates can be used for IVT, including circular plasmid DNA, (PCR-amplified) DNA fragments, and single-stranded DNA (ssDNA or ssODN).