The real time PCR (qPCR) method provides a powerful method to assess levels of particular species of DNA. When combined with reverse transcription (RT-qPCR) it is the predominate technique to measure expression of gene transcripts. While this approach is very powerful, particular care must be taken in the design of the primers to facilitate…

Recombinant protein expression and site-directed mutagenesis of target genes have demonstrated an increasing importance in the fields of molecular biology, biochemistry, biotechnology, and medicine. By using the flavodoxin of the model cyanobacterium "Anabaena" sp. PCC 7120 as a laboratory tool, we designed a comprehensive laboratory…

The content and design scheme for a readily adaptable, research-based laboratory course in chemical biology are presented. In this course, students interrogate protein structure and function using the site-specific incorporation of noncanonical amino acids by genetic code expansion. The relatively new field of genetic code expansion enables…

Disrupting a gene to determine its effect on an organism's phenotype is an indispensable tool in molecular biology. Such techniques are critical for understanding how a gene product contributes to the development and cellular identity of organisms. The explosion of genomic sequencing technologies combined with recent advances in genome-editing…

Exposure of organisms to high temperatures and various chemical and physical stressors can cause protein misfolding and aggregation. In turn, this can disrupt the functions of proteins, threatening both development and homeostasis. To overcome this, cells can initiate the highly conserved heat shock (HS) stress response pathway. In eukaryotes,…

Animal biology is fascinating for its incredible diversity in life strategies. These strategies amaze scientists and can also fire the enthusiasm of science students. One group of animals impressive in this way is the phylum "Cnidaria," containing some 10,000 invertebrate species that include jellyfish, corals, sea anemones, hydroids,…

A seven-week "gene to protein" laboratory sequence is described for an undergraduate biochemistry laboratory course. Student pairs were given the task of introducing a point mutation of their choosing into the well studied protein, enhanced green fluorescent protein (EGFP). After conducting literature searches, each student group chose the…

Molecular docking is a frequently used method in structure-based rational drug design. It is used for evaluating the complex formation of small ligands with large biomolecules, predicting the strength of the bonding forces and finding the best geometrical arrangements. The major goal of this advanced undergraduate biochemistry laboratory exercise…

Personalized medicine refers to medical care that involves genetically screening patients for their likelihood to develop various disorders. Commercial genome screening only involves identifying a consumer's genotype for a few single nucleotide polymorphisms. A phenotype (such as an illness) is greatly influenced by three factors: genes, gene…

First reported in the early 1930s, variation in the ability to taste phenylthiocarbamide (PTC) has since become one of the most widely studied of all human genetic traits. Guo and Reed (2001) provide an excellent review of work on this polymorphism prior to the identification and sequencing of the PTC gene by Kim et al. (2003), and Wooding (2006)…

A simple in class laboratory illustrating the principles of ion exchange chromatography as a bioseparation technique is described. A protein's isoelectric point as a driving force for ion exchange chromatography is easily demonstrated by using combinations of proteins with natural color or fluorescence, such as DsRed2, enhanced green fluorescent…

Noncovalent interactions and physicochemical properties of amino acids are important topics in biochemistry courses. Here, we present a computational laboratory where the capacity of each of the 20 amino acids to maintain different noncovalent interactions are used to investigate the stabilizing forces in a set of proteins coming from organisms…

Comprehensive measurement of gene expression using high-density nucleic acid arrays (i.e. microarrays) has become an important tool for investigating the molecular differences in clinical and research samples. Consequently, inclusion of discussion in biochemistry, molecular biology, or other appropriate courses of microarray technologies has…

This article describes how to establish a primary tissue culture, where cells are taken directly from an organ of a living animal. Cardiac cells are taken from chick embryos and transferred to culture dishes. These cells are not transformed and therefore have a limited life span. However, the unique characteristics of cardiac cells are maintained…

The process of diffusion is described as the movement of a substance from an area of high concentration to an area of low concentration. In this article, the authors present a novel way to introduce students to diffusion through the use of an analytical puzzle. This exercise engages students in active learning through a thought provoking exercise…