Gaussia Luciferase Plasmid Construction

Previously constructed pTarget super-luminescent Gaussia princeps luciferase (SGLuc) plasmid [6 (link)] was used as a template for the insertion of SGLuc into pMC.CMV-MCS-SV40polyA with BamHI-HF^® and EcoRI-HF^®. Ligation was performed using T4 DNA Ligase (Roche), and subsequently transformed into NEB^® 5-alpha competent E. coli (New England Biolabs) and plated on 50 µg/mL Kanamycin LB agar plates (Teknova). Selected colonies were grown in imMedia™ Growth Medium with Kanamycin (Invitrogen) overnight at 37 °C.

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Puckette M., Clark B.A., Barrera J., Neilan J.G, & Rasmussen M.V. (2023). Evaluation of DNA Vaccine Candidates against Foot-and-Mouth Disease Virus in Cattle. Vaccines, 11(2), 386.

Publication 2023

T4 DNA Ligase NEB® 5-alpha competent E. coli Kanamycin LB agar plates imMedia™ Growth Medium with Kanamycin

Agar E coli Ecori Growth medium Kanamycin Ligation Luciferase Luminescent Plasmid T4 dna ligase

Corresponding Organization : Plum Island Animal Disease Center

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Variable analysis

independent variables

Insertion of SGLuc into pMC.CMV-MCS-SV40polyA with BamHI-HF® and EcoRI-HF®

dependent variables

Successful transformation of pMC.CMV-MCS-SV40polyA with SGLuc into NEB® 5-alpha competent E. coli
Growth of selected colonies in imMedia™ Growth Medium with Kanamycin

control variables

Concentration of Kanamycin at 50 µg/mL in LB agar plates
Incubation temperature at 37 °C

positive controls

Previously constructed pTarget super-luminescent Gaussia princeps luciferase (SGLuc) plasmid

negative controls

Not explicitly mentioned

Annotations

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