Journal: bioRxiv

Article Title: The C-terminal domain of SEC-10 is fundamental for exocyst function, Spitzenkörper organization and cell morphogenesis in Neurospora crassa

doi: 10.1101/2022.01.14.475644

Figure Lengend Snippet: Tagging SEC-10 with GFP at its C-terminus disrupts growth in homokaryons, and the sec-10 knock out mutation is lethal and dominant in heterokaryons. A) Schematic view of the DNA constructs used. DNA constructs were incorporated into endogenous loci via electroporation and the strains were selected by hygromycin resistance (hph). B) Left column shows colony growth of the strains after 24 hours. Laser scanning confocal microscopy of strains stained with FM4-64 reveal vesicle distribution inside the cell. The DIC microscopy shows the difference in morphology between the different strains. The right column chart shows growth rate of the strains: wild type (orange), GFP tagged (green), and sec-10 mutants (blue). Scale bar = 10 μm.

Article Snippet: A total of 1 μg of DNA (500 ng of each DNA replacement cassette) was mixed with 1.25E8 macroconidia of the FGSC9718 strain and transferred to 0.2 cm-gap sterile electroporation cuvettes (BIO-RAD ® GenePulser XcellTM) for transformation by electroporation (1500 V, 25 μF, 600 Ohm) ( )).

Techniques: Knock-Out, Mutagenesis, Construct, Electroporation, Confocal Microscopy, Staining, Microscopy

Journal: STAR Protocols

Article Title: An in vitro CRISPR-Cas12a-mediated protocol for direct cloning of large DNA fragments

doi: 10.1016/j.xpro.2023.102435

Figure Lengend Snippet:

Article Snippet: Optional: Cut 1/4 plug using a clean blade and run the pulsed field gel electrophoresis (PFGE) with GenePulser XcellTM microbial system (Bio-Rad, Catalog# 1652662) to make sure that the target DNA fragment had been cleaved ( B).

Techniques: Virus, Recombinant, Marker, Cloning, Plasmid Preparation, Modification, Software, CRISPR, Spectrophotometry, Imaging, Electrophoresis