Bi-weekly Research Presentation for Underrepresented Minorities in STEM

Our upcoming seminar features two amazing graduate students who will be sharing with us some of their research work. Our first speaker is Gideon Ofosu Addai. Gideon graduated from KNUST in 2020 and subsequently served as a teaching assistant in the university's chemistry department. He is currently a PhD candidate at Florida State University USA, where his research focuses on the effects of organic chemistry on biological molecules in their native environments.

Our second speaker is Mainprice Essuman. He graduated from University of Cape Coast for his BSc and is currently a graduate at Southern Illinois University Edwardsville.

Speaker 1: Gideon Ofosu Addai

Topic: Chelation Chemistry and Its Importance in Improving Benzylcytosines as CLIP-Tag substrates for cellular labeling.

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Speaker 2: Mainprice Essuman

Topic: Overlap Extension PCR Cloning: A Non-standard Cloning Strategy

Abstract: Gene cloning is the production of identical copies of a particular gene or DNA sequence using genetic engineering techniques and has wide application in biomedical research. The existing molecular cloning methods mostly involve the use of restriction enzymes, and are often limited by the availability of suitable restriction sites. It is still a challenge for simultaneous cloning of multiple fragments into different sites of a single vector. Here, I describe overlap extension PCR (OEP) cloning, a straightforward, efficient, and reliable way to clone an insert of choice into a plasmid of choice without restriction endonucleases or T4 DNA ligase. In this technique, chimeric primers containing plasmid sequence at the 5′ ends and insert sequence at the 3′ ends are used to PCR-amplify insertion sequences of various sizes. These inserts are employed as mega-primers in a second PCR with a circular plasmid template. The original plasmid templates are then destroyed using DpnI, and the OEP products used to transform competent Escherichia coli cells. Phusion DNA polymerase is used for the amplification and fusion reactions making the reactions easy to monitor and optimize. Findings of independent studies shows that products of OEP cloning are dependent on PCR cycles, polymerase used, and length of inserts. An improved OEP cloning technique improves efficiency of OEP, allows for cloning of multiple fragments at the same time, and can be used for large fragments. OEP cloning presents an efficient alternative to traditional methods, enabling the cloning of multiple fragments simultaneously without the need for restriction enzymes or ligases.