Biochem 14.2 Blotting Techniques and Methods Quiz

<p>Salmon sperm DNA (C)</p> Signup and view all the answers

<p>They are antibodies that specifically recognize the target protein (B)</p> Signup and view all the answers

<p>Minimizes background noise (D)</p> Signup and view all the answers

<p>All of the above (D)</p> Signup and view all the answers

<p>Probes are designed to bind specifically to target molecules only (D)</p> Signup and view all the answers

<p>They are quicker and less labor-intensive. (D)</p> Signup and view all the answers

<p>Denaturing the DNA to single strands. (A)</p> Signup and view all the answers

<p>It denatures the double-stranded DNA. (C)</p> Signup and view all the answers

<p>Upward capillary transfer. (A)</p> Signup and view all the answers

<p>Dot blots do not involve electrophoresis. (A)</p> Signup and view all the answers

<p>It must be denatured to single strands. (A)</p> Signup and view all the answers

<p>They provide less resolution and molecular size data. (A)</p> Signup and view all the answers

<p>In native conditions to keep DNA double-stranded. (C)</p> Signup and view all the answers

<p>To prevent structured regions from affecting migration (B)</p> Signup and view all the answers

<p>Analysis of various RNA subtypes (D)</p> Signup and view all the answers

<p>Electroblotting (D)</p> Signup and view all the answers

<p>To maintain directional transfer of proteins (C)</p> Signup and view all the answers

<p>They need additional treatment to ensure migration direction (A)</p> Signup and view all the answers

<p>It generates more heat during the process (D)</p> Signup and view all the answers

<p>To bind proteins as they leave the gel (A)</p> Signup and view all the answers

<p>cDNA (C)</p> Signup and view all the answers

<p>To immobilize DNA after it rises with the buffer (C)</p> Signup and view all the answers

<p>Block the membrane (C)</p> Signup and view all the answers

<p>It hydrolyzes RNA (A)</p> Signup and view all the answers

<p>The annealing of an oligonucleotide probe to target DNA (D)</p> Signup and view all the answers

<p>Gene duplication or deletion testing (D)</p> Signup and view all the answers

<p>RNA requires a chemical denaturant due to instability (C)</p> Signup and view all the answers

<p>The hybridized target-probe pair can be visualized (B)</p> Signup and view all the answers

<p>Northern blots focus on RNA while Southern blots focus on DNA (A)</p> Signup and view all the answers

<p>To allow for visualization and detection of the probe (A)</p> Signup and view all the answers

<p>Via radioactive decay that interacts with radiographic film (B)</p> Signup and view all the answers

<p>To visualize the decay of radioactive labels on film (A)</p> Signup and view all the answers

<p>They do not disrupt the chemical properties of probe molecules (D)</p> Signup and view all the answers

<p>They require specialized digital cameras for detection (A)</p> Signup and view all the answers

<p>It helps indicate relative changes without precise quantities (B)</p> Signup and view all the answers

<p>32P (D)</p> Signup and view all the answers

<p>To assess changes in the expression of target molecules (C)</p> Signup and view all the answers

<p>To prevent non-specific binding (C)</p> Signup and view all the answers

<p>They reduce the cost and improve signal detection (D)</p> Signup and view all the answers

<p>Enzymes or fluorescent molecules (A)</p> Signup and view all the answers

<p>By binding multiple times to primary antibodies (A)</p> Signup and view all the answers

<p>They specifically recognize target proteins. (B)</p> Signup and view all the answers

<p>They block non-specific binding sites on the membrane. (D)</p> Signup and view all the answers

<p>Multiple techniques including immunohistochemistry and ELISAs (C)</p> Signup and view all the answers

<p>Chemiluminescent or chromogenic reactions (B)</p> Signup and view all the answers

Nitrocellulose and polyvinylidene fluoride (PVDF) membranes are commonly used in protein analysis. They bind proteins through a combination of hydrophobic and electrostatic interactions.

Capillary transfer and electroblotting are two methods used to move samples from an electrophoretic gel to a membrane.

Probes bind to the target molecule of interest on the membrane, allowing for its visualization.

Blotting membranes contain nonspecific binding sites that can lead to background noise. Blocking solutions are used to saturate these sites, minimizing false signals and increasing clarity.

Southern blots typically use single-stranded DNA primers to bind to specific DNA sequences on the membrane.

Western blots typically use antibodies that specifically recognize the target protein.

Salmon sperm DNA is often used to block nonspecific binding in Southern and Northern blotting.

Bovine serum albumin (BSA) and fat-free cow's milk are often used to block nonspecific binding in Western blotting.

A method of labeling molecules using radioactive isotopes.

A type of film that detects radioactive decay, showing the location of labeled molecules on a membrane.

A visual representation of the signals from the radioactive labels on a blotting membrane.

A technique for detecting molecules based on their ability to emit light when exposed to a specific wavelength.

A technique for detecting molecules based on their ability to absorb light at a specific wavelength and emit light at a different wavelength.

A method for comparing the amount of a specific molecule across samples using visualization of bands.

A reference molecule added to ensure that different samples are loaded and treated equally during blotting experiments.

A molecule that binds specifically to the target molecule and can be detected by a label.

A technique used to detect specific DNA sequences in a sample, where DNA is transferred from an agarose gel to a membrane, then hybridized with a labeled probe.

A technique used to detect specific RNA sequences in a sample, where RNA is transferred from an agarose gel to a membrane, then hybridized with a labeled probe.

A process where a single-stranded oligonucleotide probe, complementary to a specific sequence in the target DNA or RNA, binds to the target nucleic acid.

The process of blocking nonspecific binding sites on a blotting membrane. This is done to minimize background noise and increase the clarity of the signal.

A solution used to block nonspecific binding on blotting membranes. Common reagents include salmon sperm DNA, bovine serum albumin (BSA), or fat-free milk.

A chemical denaturant, such as formaldehyde, is used to break down secondary structures in RNA, allowing for more efficient transfer to a membrane.

Unlike Southern blots, which detect DNA, Northern blots target RNA. The naming system is based on the direction of the target nucleic acid.

Alkaline denaturation, which hydrolyzes RNA, cannot be used in Northern blotting because RNA is more reactive than DNA.

A technique used to analyze RNA expression and sequence by separating RNA molecules based on size, transferring them to a membrane, and detecting specific RNA sequences using labeled probes.

A technique used to analyze protein expression by separating proteins based on size, transferring them to a membrane, and detecting specific proteins using labeled antibodies.

Formaldehyde is used during northern blotting to maintain the structure and prevent RNA folding. It acts like SDS in SDS-PAGE, causing denaturation.

A membrane used in blotting techniques to bind and retain the separated molecules, facilitating further analysis.

A method of transferring proteins from a gel to a membrane using an electric field.

A method of visualizing specific molecules on a blotting membrane by using probes that are labeled with a detectable signal.

A type of blocking solution containing proteins like bovine serum albumin (BSA) or casein, used to prevent nonspecific binding of probes to the membrane.

A type of blotting technique that avoids gel electrophoresis, instead directly spotting samples onto a membrane.

The process of transferring DNA fragments from an agarose gel to a blotting membrane.

A method of transferring DNA fragments from an agarose gel to a membrane using capillary action and buffer.

In Southern blotting, the DNA is denatured into single strands before transfer to the membrane.

A specific sequence of DNA or RNA used to bind to a complementary sequence on a membrane in Southern and Northern blots.

The step after transfer where the membrane is incubated with a blocking solution to prevent nonspecific binding of probes.

The process of visualizing the target molecule that has been bound to the membrane with a probe.

Using a secondary antibody that recognizes and binds to the primary antibody, which directly binds to the target protein.

The use of secondary antibodies allows the same secondary antibody to be used with different primary antibodies from the same species, reducing the need for expensive and time-consuming labeling of each primary antibody.

Secondary antibodies often bind to multiple epitopes on the primary antibody, increasing the signal strength. This is due to their dimeric nature and polyclonal composition.

The enzyme horseradish peroxidase (HRP) is commonly used to catalyze a chemiluminescent or chromogenic reaction. The reaction produces a visible signal for detection.

In a Western blot, a fluorophore, a molecule that absorbs light at a specific wavelength and emits light at a different wavelength, is linked to an antibody.

This technique measures the amount of protein in a sample by comparing the intensity of the band on the Western blot with a known standard.

A loading control is a reference protein added to ensure equal loading and treatment of different samples in the Western blot.

In Western blotting, an antibody is employed as a probe to specifically bind to the protein of interest on the membrane. This allows for its visualization and detection.