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Resources and Experiments for the HOSA Biotechnology Event

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A key component of the HOSA biotechnology event is development of technical skills that take your students out of the books and into the laboratory.  That is why we are happy to provide experiments and technical skills tutorials to aid in your training for both components of this competitive event.

Laboratory Basics:

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Micropipetting:  

Pipettes are essential laboratory tools used by scientists to measure and manipulate liquids. They range from simple eye dropper-like devices with squeezable bulbs to advanced robotic systems capable of precise volume dispensing. Micropipettes, in particular, play a crucial role in accurately measuring small volumes, ensuring successful and reproducible experiments.

In biotechnology labs, where reactions involve microliter volumes, scientists rely on piston displacement micropipettes. The accuracy of pipetting is of utmost importance in these experiments, as even minor inconsistencies like air bubbles or extra droplets can disrupt reaction proportions. Therefore, to optimize your students’ laboratory results, it is critical to make sure that they have mastered the technical skill of micropipetting.

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Restriction Enzymes:

One of the most significant discoveries in molecular biology belongs to a group of enzymes called restriction endonucleases, also referred to as restriction enzymes. These remarkable enzymes function like molecular scissors by precisely cleaving through the sugar-phosphate DNA backbone based on the specific sequence of nucleotides.  The immense utility of restriction enzymes revolutionized numerous scientific endeavors, including molecular cloning, DNA mapping, sequencing, and a wide array of genome-wide studies.  This discovery launched the era of modern molecular biotechnology.

Structure of EcoRI restriction enzyme homodimer (cyan and green) bound to DNA (tan). 

Agarose Gel Electrophoresis and Gel Interpretation:

Agarose gel electrophoresis is a biotechnology technique that uses electricity and a porous gel matrix to separate mixtures of molecules based on their charge, shape, and size. It is commonly used to separate dyes, proteins, and nucleic acids like DNA and RNA.  In particular, it is especially useful in analyzing mixtures of DNA fragments generated by restriction enzyme digestion. During agarose gel electrophoresis, these DNA fragments are separated into distinct bands according to their respective sizes.

Bacterial Transformation and Transformation Efficiency:

In nature, non-essential genes are found on small circular pieces of double-stranded DNA known as plasmids. These plasmids facilitate the exchange of beneficial genes among bacteria and can replicate independently from the cell’s chromosomal DNA.  In the lab, we can modify plasmids by introducing genes from various sources. When these engineered plasmids are introduced into bacteria, they transform the bacteria into living factories capable of producing valuable substances like medications, vitamins, and insulin, which is used to treat diabetes.

Since E. coli are not naturally competent, we need to force them to take up plasmid DNA in the lab. This can be done with electricity, in a process called electroporation, or through physical means in a heat shock.  In a heat shock transformation, the cells are treated with calcium chloride to make them “competent”.  DNA is added to the cells before they are moved quickly between two very different temperatures. It is believed that the combination of calcium chloride and the rapid change in temperature changes the permeability of the cell wall and membrane, allowing DNA molecules to enter the cell.  

In practice, transformation is highly inefficient—only one in every 10,000 cells successfully incorporates the plasmid DNA. However, since many cells are used in a transformation experiment (about a billion cells), only a few cells must be transformed to achieve a positive outcome. We can use the data from our experiment to determine how well our transformation worked.  

Qualitative ELISA:  

The Enzyme-Linked Immunosorbent Assay, or ELISA, is a highly sensitive test that uses antibodies to detect the presence of specific molecules within a complex sample.  is an invaluable tool for researchers, as it can identify even trace amounts of antigens in biological samples, making it ideal for pathogen and allergen detection, among other applications. The fundamental principle of ELISA revolves around the use of antibodies to identify antigens present in the samples. With the capability to generate antibodies for a wide range of molecules, ELISA has emerged as a versatile laboratory test.

Polymerase Chain Reaction (PCR):

In the early 1980’s, Kary Mullis developed a technique that replicated DNA in vitro using short, synthetic DNA oligonucleotides designed to target a specific sequence (known as primers) and DNA Polymerase I. In a process similar to replication in a cell’s nucleus, the primers would bind to the DNA, directing polymerase to copy the gene sequence. However, after the initial elongation, the sample was heated to denature the newly-formed DNA duplex, then cooled to allow primer binding and extension to happen again. Each time the sample cycled through the different temperatures, the amount of DNA doubled. By repeating this cycle of heating and cooling many times, billions of copies of a specific DNA sequence were produced in a matter of minutes. This simple cycle – anneal, extend, denature – is the basis of the polymerase chain reaction.  Because of its ease of use and its ability to rapidly amplify DNA, PCR has become indispensable in the medical and life sciences lab.  

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