title: pELMO, an optimised in-house cloning vector.
abstract: DNA cloning is an essential tool regarding DNA recombinant technology as it allows the replication of foreign DNA fragments within a cell. pELMO was here constructed as an in-house cloning vector for rapid and low-cost PCR product propagation; it is an optimally designed vector containing the ccdB killer gene from the pDONR 221 plasmid, cloned into the pUC18 vector's multiple cloning site (Thermo Scientific). The ccdB killer gene has a cleavage site (CCC/GGG) for the SmaI restriction enzyme which is used for vector linearisation and cloning blunt-ended products. pELMO transformation efficiency was evaluated with different sized inserts and its cloning efficiency was compared to that of the pGEM-T Easy commercial vector. The highest pELMO transformation efficiency was observed for ~500 bp DNA fragments; pELMO vector had higher cloning efficiency for all insert sizes tested. In-house and commercial vector cloned insert reads after sequencing were similar thus highlighting that sequencing primers were designed and localised appropriately. pELMO is thus proposed as a practical alternative for in-house cloning of PCR products in molecular biology laboratories.
authors.affiliation: Molecular Biology and Immunology Department, Fundación Instituto de Inmunología de Colombia, Cra. 50 # 26-20, Bogotá, Colombia.
authors.forename: Andrea E
authors.initials: AE
authors.lastname: Ramos
authors.affiliation: Molecular Biology and Immunology Department, Fundación Instituto de Inmunología de Colombia, Cra. 50 # 26-20, Bogotá, Colombia.
authors.forename: Marina
authors.initials: M
authors.lastname: Muñoz
authors.affiliation: Molecular Biology and Immunology Department, Fundación Instituto de Inmunología de Colombia, Cra. 50 # 26-20, Bogotá, Colombia.|PhD Programme in Biomedical and Biological Sciences, Universidad del Rosario, Bogotá, Colombia.
authors.forename: Darwin A
authors.initials: DA
authors.lastname: Moreno-Pérez
authors.affiliation: Molecular Biology and Immunology Department, Fundación Instituto de Inmunología de Colombia, Cra. 50 # 26-20, Bogotá, Colombia. mapatarr.fidic@gmail.com.|School of Medicine and Health Sciences, Universidad del Rosario, Bogotá, Colombia. mapatarr.fidic@gmail.com.
authors.forename: Manuel A
authors.initials: MA
authors.lastname: Patarroyo
keywords: Blunt-ended; Cloning vector; PCR cloning; Recombinant DNA technology; ccdB killer gene