Journal: Journal of genome editing and regulation

Article Title: Generating a New sgRNA Vector, pGL3-U6-sgRNA-PGK-mRFP-T2A-PuroR, to Improve Base Editing

doi: 10.32371/jger/246146

Figure Lengend Snippet: A) Schematic diagram of NheI-based RFLP analysis of potential Munc13-1null clones. B). A 799bp Munc13-1 DNA fragment containing the editing site was amplified from individual clones using One-Taq DNA polymerase. Following NheI digestion and subsequent agarose (1.5%) gel electrophoresis, monoallelic mutants were distinguished from the WT.

Article Snippet: From plasmid MCS1-EF1α-RFP-T2A-Puro-pA-MCS2, the mRFP cDNA and the downstream T2A linker were amplified in a 50 μl touchdown PCR ( ) that contains 150 ng of the template DNA, 200 μM dNTPs, 5 μl of 10x pfu buffer, 5 units of Phusion DNA polymerase (G-Biosciences), and 200 nM forward and reverse primers (mRFPF and mRFPR).

Techniques: Clone Assay, Amplification, Nucleic Acid Electrophoresis