Biomedicine Methods Exam

The process of inserting a specific DNA sequence into a vector. This vector can then be used to introduce the sequence into a host cell for various applications.

A process of visualizing and analyzing protein expression levels in samples using antibodies. It helps identify and quantify specific proteins.

A technique that involves reducing the expression of a specific gene using a short RNA molecule that binds to the gene's mRNA transcript, preventing protein translation.

A protein that plays a crucial role in regulating cell growth, survival, and metabolism. Its phosphorylation state indicates its activation and involvement in various cellular processes.

The phenomenon where distinct antibodies that recognize specific proteins can exhibit variability in their binding efficiency, resulting in differing signal intensities during Western Blot analysis. It's essential to standardize and validate antibody performance for reliable experimental results.

A technique used to quantify protein levels in a sample by analyzing the intensity of protein bands on a Western blot membrane.

The process of using a blocking solution to prevent non-specific binding of antibodies to the membrane in a Western blot experiment.

The process of transferring proteins from a gel to a membrane in a Western blot experiment.

In Western blotting, this indicates the degree of precision and reliability in determining protein levels.

A control sample that is treated identically to the experimental samples, except it lacks the factor being investigated. It helps to determine if any observed changes in the experimental samples are due to the specific factor being studied.

A control sample used in Western blotting to ensure that the amount of protein loaded into each lane is equal. It helps to adjust for any variations in loading and ensures a fair comparison between samples.

A type of antibody that specifically binds to the primary antibody. It is typically conjugated to an enzyme or fluorescent molecule to enable detection.

Non-specific binding of antibodies to the membrane, resulting in a background signal that can obscure the specific signal of the target protein.

The practice of manipulating images in a scientific publication in a way that misrepresents the data.

Making small adjustments to data in a way that supports a researcher's hypothesis, even if the adjustments are not fully justified by the data.

A plasmid vector that has been engineered to enhance its efficiency and safety. It typically incorporates features like multiple cloning sites (MCS) for easy gene insertion and stronger promoters for increased gene expression. It may also include elements for improved stability or reduced immunogenicity.

A virus that has been modified to carry and deliver genetic material into cells. Viral vectors are commonly used in gene therapy and research to introduce genes into cells of interest.

A viral vector that has been further developed for enhanced safety and efficiency. It may incorporate features like capsid modifications for cell-specific targeting, better promoters for increased expression, and reduced immunogenicity.

A type of plasmid vector that carries a gene for resistance to the antibiotic neomycin. This gene allows researchers to easily select for cells that have successfully taken up and integrated the plasmid.

A way to obtain functional eukaryotic proteins in a prokaryotic system. By using a eukaryotic cDNA instead of the whole gene, only the coding sequence is included, avoiding issues with prokaryotes inability to process introns.

A type of gel used in electrophoresis, with a high percentage of acrylamide (typically 7% or higher) to separate large proteins with molecular masses of 200kDa or higher.

To study glucose regulation, a researcher needs the insulin protein. Cloning the insulin gene and expressing it in a suitable host allows for production of the desired protein.

Recombinant insulin can be purified to be used in experiments related to glucose regulation. Purifying the molecule isolates the desired protein, allowing for accurate and controlled experimentation.

The protein samples would not sink to the bottom of the gel, and they would not move through the gel matrix effectively. This is because glycerol is used to add density to the samples, allowing them to sink into the wells of the gel.

A higher acrylamide percentage gel is used for separating smaller proteins. 20% acrylamide would be the most suitable for a protein of 200kDa.

It normalizes the signal, allowing you to compare the amount of target protein to a known, stable reference. This helps you eliminate variability in protein levels caused by factors unrelated to your experiment.

Two unique restriction sites ensure that the vector won't recircularize without the inserted gene. This prevents the plasmid from closing up without the gene of interest.

A viral vector is often the preferred choice because it efficiently integrates the gene into the host cell's genome, ensuring stable expression of the gene over multiple generations. Some popular examples of viral vectors include lentiviral vectors and retroviral vectors.

SDS-PAGE is a technique used to separate proteins based on their molecular weight. The negatively charged SDS molecules bind to and denature proteins, giving them a uniform negative charge. When applied to an electric field, proteins migrate through a gel matrix at a speed proportional to their size.

Restriction enzymes cut DNA at specific sequences called restriction sites, acting like molecular scissors. This process is used in genetic engineering to manipulate DNA.

A plasmid is a small, circular DNA molecule that can be used as a vector to carry genes into cells. It contains an origin of replication for self-replication, a selectable marker gene for identification of transformed cells, and regions for inserting foreign DNA.

Transfection is the process of delivering genetic material into eukaryotic cells. This can be achieved through various methods, including chemical transfection, electroporation, and viral vectors.

A control is crucial for any experiment to ensure that observed changes are caused by the experimental variable and not other factors. A loading control provides a baseline for comparison, helping determine if changes in the target protein are real or due to other factors.