Qiongqiong Zhou

Investigating the Physico-Chemical Properties of Amino Acids and Their Analysis by Thin Layer Chromatography

In this laboratory, you study the solubility and thin layer chromatography (TLC) behavior of representative amino acids. Four amino acids—proline and leucine (nonpolar), cysteine (polar), and lysine (ionic)—are used as amino acid standards to identify an unknown amino acid based on interpretation of the TLC pattern. To do that, you employ a matrix design to assess solubility and then separate these amino acids on a TLC plate and stain them with ninhydrin. Based on the Rf values and colors of the stained amino acid standards, you infer what the unknown is.

Preparing Buffers at a Specific Molarity and pH

In this laboratory, you learn how to correctly make three of the most common buffers used today in biochemistry or cell and molecular biology. The buffers Tris-acetate-EDTA, phosphate buffered saline, and Tris-glycine-SDS are abbreviated TAE, PBS and TGS, respectively.

An Introduction to Basic Math and Operations in the Biomolecular Laboratory

In this laboratory experiment, you review some of the basic equipment commonly used in biomolecular science, such as the mass balance and pipettes. You refamiliarize yourself with the proper use and handling of this equipment and some of the typical math and conversions you must use to successfully perform work in the lab, such as mass, volume, and concentration. Finally, you review some of the simple statistical parameters (range, average, standard deviation, and student t-test) used in analyzing your data.

Studying Cell-like Structures with Liposome, DNA, and Protein

In this laboratory experiment, you prepare a simple liposome containing a phospholipid surrounding internalized DNA. The lipid layer and DNA are fluorescently labeled or stained and then analyzed by using fluorescence microscopy.

Investigating Protein: Nucleic Acid Interactions by Electrophoretic Mobility Shift Assay (EMSA)

In this laboratory experiment, you investigate a typical protein:nucleic acid interaction, using the model protein protamine and plasmid DNA (pDNA). When the DNA is bound by protein, its migration pattern and distance through the gel will vary depending on how much protein is bound to it. This demonstrates the classic technique called electrophoretic mobility shift assay (EMSA) that is used for analyzing the interactions between nucleic acid and protein.

Preparation of a Fluorescently Labeled Liposome and Its Analysis by Fluorescence Microscopy

In this laboratory experiment, you formulate a simple liposome composed of a triacylglyceride (tripalmitin) and cholesterol. A small amount of fluorescently conjugated cholesterol is incorporated to impart fluorescence characteristics upon the liposome. The liposome is then viewed by using fluorescence microscopy.

Rapid Purification, Gel Electrophoresis, and Enzyme Activity Assay of the Luciferase Enzyme from Fireflies

In this experiment, you perform a rapid purification and analysis of the protein enzyme Luciferase (Luc). You extract the enzyme from the tissue of the firefly tail by sonicating in buffer. The dissolved proteins are then separated from the rest of the tissue debris by centrifugation, and then the proteins in the supernatant are precipitated by the addition of acetone. The proteins are resuspended and dissolved in PBS buffer and analyzed for purity by gel electrophoresis and enzyme activity by bioluminescent assay.

Hexokinase and G6PDH Catalyzed Reactions of Glucose Measurement

Abstract The purpose of this lab is for you to gain an understanding of how glucose concentration can be accurately determined based on your hands-on experience. Here, you use glucose-specific enzyme to measure the glucose concentrations in cell culture media and juice. Enzymatic glucose measurement is more accurate than traditional chemicalreducing methods, particularly when other reducing chemicals are present in the samples. In this lab, you are asked to generate a standard curve and calculate your sample concentrations based on the standard curve and your sample measurements. The learning objectives are to 1. Learn how to make a standard curve 2. Learn and understand how to dilute samples for an accurate measurement 3. Understand how to use the standard curve to calculate sample concentration 4. Understand why sample absorptions need to be corrected by subtracting various backgrounds.

Polymerase Chain Reaction (PCR)

Abstract In this lab, you gain hands-on experience in preparing a PCR reaction. You get additional experience in making dilutions and accurately transferring microliter volumes, which will critically affect the outcome of your experiment. You also work as part of a team to pour an agarose gel and analyze your PCR products by electrophoresis. In addition, you get a chance to learn how to use the National Center for Biotechnology Information (NCBI) and the Basic Local Alignment Search Tool (BLAST) to find out the size and the sequence of your amplified gene. The learning objectives are to 1. Put into practice a basic PCR procedure 2. Improve mathematical, analytical, and technical skills 3. Become familiar with NCBI

Experiment 8 – Qualitative Analysis of the Degradation of RNA via Ribonuclease A versus B

RNA has traditionally been somewhat of the “black box” of molecular biology. Today we have a fairly deep understanding of genomics and proteomics, but the myriad of RNA structures and functions RNA serves in cells and tissues in regulating gene expression has gone vastly underappreciated. There are many RNases present in cells and tissues that direct the cleavage and regulate the half-life of RNA—one wellstudied example being RNase A and its glycosylated version, RNase B. In this laboratory experiment, you gain hands-on experience working with RNA by comparing its degradation by RNase A and B and investigating the kinetics of its reaction.