biotin receiver strands biotin handle invader agaacgcctatcactacc scaffold production helper plasmid hp17 ko7  (Addgene inc)

Journal: The EMBO Journal

Article Title: PUFFFIN: an ultra-bright, customisable, single-plasmid system for labelling cell neighbourhoods

doi: 10.1038/s44318-024-00154-w

Figure Lengend Snippet: Reagents and tools table

Article Snippet: EMMA assemblies of parts into the YCe3736_HC_Amp_ccdB_receiver vector (a kind gift from Prof Steven Pollard, also available on Addgene: #100637) were carried out using the NEBridge Golden Gate Assembly Kit (BsmBI-v2) (NEB, #E1602L), using 75 ng of each part and 2 μl of enzyme mix in a 20 μl reaction, performing 60 cycles of digestion (42 °C, 5 min) and ligation (16 °C, 5 min), followed by heat inactivation of enzymes (60 °C, 5 min).

Techniques: Subcloning, Recombinant, Plasmid Preparation, Modification, Saline, Produced, Transfection, Cell Culture, SYBR Green Assay, Staining, PCR Cloning, Software, High Content Screening, Microscopy