RNase one gene isolation, expression, and affinity purification models research experimental progression and culminates with guided inquiry‐based experiments

Abstract

This new biochemistry laboratory course moves through a progression of experiments that generates a platform for guided inquiry-based experiments. RNase One gene is isolated from prokaryotic genomic DNA, expressed as a tagged protein, affinity purified, and tested for activity and substrate specificity. Student pairs present detailed explanations of materials and methods and the semester culminates in a poster session. Experimental plans take into account the expense and time required to move from gene isolation to enzyme assays. This combination of instructor-guided and student-designed experiments is a manageable foray into guided inquiry-based learning in a biochemistry laboratory course, while providing a cohesive story and context for individual experiments.Keywords: guided-inquiry, RNase one, experimental design.Students typically retain only 20% of the content of any given course [1,2], and most college students, even high achieving students, continue to think in the same ways they thought prior to taking their courses [3]. These results give pause for thought to even the most gifted teacher. The reasons for such low content retention output are various but include the failure to create learning environments that help students learn how to think for themselves and learn on their own, assess stated learning objectives other than rote memorization, and actively construct their own learning [1,2]. Active learning is particularly important for achieving transferrable, critical thinking, communication, and other ''soft'' skills that students need for living and working in a globalized world [4].Inquiry-based learning environments are more effective than traditional lecture or recipe style laboratory courses. Sixth and seventh grade students who learned physics by way of inquiry-based pedagogies outperformed eleventh and twelfth grade students who learned physics in traditional lecture formats on assessments of their grasp of physics concepts [5]. If learning is about changing the way students think, that is, changing the way they approach problems, solve problems, and make decisions in a complex, globalized world, then new learning pedagogies seem to be in order. To move toward more active, constructivist learning pedagogical models, BIO2010 [6] calls for the way scientists think through a research question to serve as a model for student learning. The inquiry based biochemistry lab curriculum described below aims to begin this kind of pedagogical change.Moving from ''cookbook'' style laboratory classes to inquiry-based or guided-inquiry based experiences can provide challenges for a faculty member instigating this change. Three such challenges are covering a required set of techniques, acquiring equipment and supplies to support a variety of experiments, and guiding an unstructured experience [7][8][9]. The semester described here combines the research process, student delivered laboratory materials and methods, two guided inquiry-based experiments, and a culminating final poster session to provide context to...

Implementation of a Collaborative Series of Classroom-Based Undergraduate Research Experiences Spanning Chemical Biology, Biochemistry, and Neurobiology

“…For our CURE design, faculty research directions had to be realigned to fit more closely to a standard laboratory session, to match with the budgetary framework available for the courses, and to maximize student success on course projects (Shortlidge et al ., 2016). One approach we used to overcome these hurdles and to minimize initial faculty input was to design each individual CURE laboratory with a parallel structure and framework to previous CURE laboratories, for example, protein structure manipulation by mutagenesis and parallel synthesis for library construction (Bailey, 2009; Knutson et al ., 2010a; Scott et al ., 2015). These overall structures were tweaked to incorporate our specific experimental methods and realigned around the central themes of mycobacterial hydrolases and neural signaling.…”

“…The laboratory met once per week for 4 hours and also fulfilled university requirements for a writing-intensive course. The experimental design for BIOCHEM paralleled similar semester-long experiments at other institutions relating the structure and function of a protein using site-directed mutagenesis, but fused this overall course design with the larger scientific goal of characterizing important mycobacterial serine hydrolases (Bailey, 2009; Knutson et al ., 2010a; Kreiling et al ., 2011). …”

Special‐study modules in a problem‐based learning medical curriculum: An innovative laboratory research practice suppporting introduction to research methodology in the undergraduate curriculum

“…A significant perspective of this work, which merits further discussion, is the practical training aspect, which is integrated with the research perspective. There is a growing emphasis on the planning, methodology, and evaluation of laboratory practicals as components of undergraduate education [18, 20, 21]. Recently, Parra et al .…”

A Review of Biochemistry Education Research

“…125 A progression of guided-inquiry experiments that combined instructor-guided and student-designed experiments was shown to help students reason through experimental procedures and findings and articulate clear research questions. 126 A pretest/post-test design applied to a course that incorporated inquiry-based learning activities within the context of PCR techniques found significant improvements in students' perceptions of their knowledge acquisition and their confidence in trouble-shooting and solving problems in the laboratory. 127 A laboratory setting that involved guiding students' learning using an inquiry-based teaching approach based on an instructor-created template that provided frequent feedback on students' work in a collaborative environment led to improvements in both students' satisfaction with the introductory biochemistry course and in the quality of the students' work.…”

“…A mixed-method study of inquiry-based laboratory techniques in which students gradually “took over” the experiment, starting from step-by-step instruction and working toward a stage in which they were involved in designing laboratory procedures, was found to improve students’ skills related to autonomy and ownership. , A study based on a similar curriculum design found improvements in students’ laboratory skills, their ability to make choices in the design of experiments, and their critical thinking skills . A progression of guided-inquiry experiments that combined instructor-guided and student-designed experiments was shown to help students reason through experimental procedures and findings and articulate clear research questions …”