dsdna donor  (Macrogen)

Journal: STAR Protocols

Article Title: Multiplexed CRISPR-Cas9 protocol for large transgene integration into the Schistosoma mansoni genome

doi: 10.1016/j.xpro.2024.102886

Figure Lengend Snippet: Schematic diagram of the long double stranded DNA donor Donor contains 5′phosphorothioate modification at the first 5 nt of both DNA strands. Plasmid DNA encoding gene cassette of an ubiquitin promoter driven EGFP with its terminator (green line) flanked on both sides by homology arms (HA) (yellow lines). A pair of 5′-phosphorothioated (P) primers (blue arrows) is used to amplify the dsDNA. PCR products are purified using a DNA column purification kit and inspected by agarose gel electrophoresis comparing with non-purified PCR (primer dimer contamination) amplicon. Purified DNA from the PCR, not from agarose gel, serves as the donor template for HDR.

Article Snippet: Assess the concentrated and purified dsDNA donor by spectrophotometry (NanoDrop, Thermo Fisher).

Techniques: Modification, Plasmid Preparation, Purification, Agarose Gel Electrophoresis, Amplification

Journal: STAR Protocols

Article Title: Multiplexed CRISPR-Cas9 protocol for large transgene integration into the Schistosoma mansoni genome

doi: 10.1016/j.xpro.2024.102886

Figure Lengend Snippet: PCR reaction master mix for long dsDNA donor preparation

Article Snippet: Assess the concentrated and purified dsDNA donor by spectrophotometry (NanoDrop, Thermo Fisher).

Techniques: Plasmid Preparation