Journal: STAR Protocols

Article Title: Protocol to identify the core gene supported by an essential gene in E. coli bacteria using a genome-wide suppressor screen

doi: 10.1016/j.xpro.2023.102196

Figure Lengend Snippet:

Article Snippet: Reagent Final concentration Amount pKDsgRNA- ack (100 ng/μL) (Addgene, #62654) 2 ng/μL 1 μL Primer pKDsgRNA F (10 μM) 200 nM 0.5 μL Primer pKDsgRNA R (10 μM) 200 nM 0.5 μL dNTPs (10 mM each) 250 μM each 0.625 μL 5× Phusion Buffer 1× 5.0 μL Phusion HF DNA Polymerase 0.04 U/μL 0.5 μL ddH 2 O N/A 16.9 μL Total N/A 25.0 μL Open in a separate window PCR setup. b. Analyze the PCR product on a 1.0% agarose gel.

Techniques: Virus, Recombinant, DNA Library Preparation, DNA Purification

Journal: STAR Protocols

Article Title: Protocol to identify the core gene supported by an essential gene in E. coli bacteria using a genome-wide suppressor screen

doi: 10.1016/j.xpro.2023.102196

Figure Lengend Snippet: PCR setup

Article Snippet: Reagent Final concentration Amount pKDsgRNA- ack (100 ng/μL) (Addgene, #62654) 2 ng/μL 1 μL Primer pKDsgRNA F (10 μM) 200 nM 0.5 μL Primer pKDsgRNA R (10 μM) 200 nM 0.5 μL dNTPs (10 mM each) 250 μM each 0.625 μL 5× Phusion Buffer 1× 5.0 μL Phusion HF DNA Polymerase 0.04 U/μL 0.5 μL ddH 2 O N/A 16.9 μL Total N/A 25.0 μL Open in a separate window PCR setup. b. Analyze the PCR product on a 1.0% agarose gel.

Techniques: Concentration Assay