All GRE Subject Test: Biochemistry, Cell, and Molecular Biology Resources
Example Questions
Example Question #31 : Gre Subject Test: Biochemistry, Cell, And Molecular Biology
When performing whole-mount immunofluorescence of a specimen, background staining is often a concern with custom generated antibodies. Which of the following is not a way in which to increase specificity of the custom antibody?
Generate the custom antibody against purified protein rather than a predictive peptide
Affinity purify the custom antisera
Determine optimal antibody dilution for the application by performing serial dilutions
Determine optimal blocking buffer components and concentrations
None of the other answers
None of the other answers
The correct answer is none of the other answers. To increase specificity of the generated antibody, it is beneficial to expose the animal (in which the antibody will be made) to the entire purified protein, rather than a short synthesized predictive peptide. Moreover, affinity purifying the antibody by exposing the bled antisera to the purified protein and separated the bound antibody-protein fraction from antibody and protein alone will increase specificity. Following antibody generation, determining the optimal dilution of the antibody as well as appropriate blocking conditions to compete out non-specific binding with the specimen are necessary.
Example Question #32 : Gre Subject Test: Biochemistry, Cell, And Molecular Biology
When is indirect immunofluorescence of sectioned specimens beneficial over whole-mounted specimens?
Sectioning if preferred when using RNA probes, whereas whole-mount is solely for antibody staining
Sectioning is preferred if the whole-mount is too large in the X plane to resolve by confocal microscopy
Sectioning is preferred if the whole-mount is too large in the Z plane to resolve by confocal microscopy
Sectioning is preferred if the whole-mount is too large in the Y plane to resolve by confocal microscopy
Whole-mount staining is only for cryo-preserved specimens
Sectioning is preferred if the whole-mount is too large in the Z plane to resolve by confocal microscopy
The correct answer is sectioning is preferred if the whole-mount is too large in the Z plane to resolve by confocal microscopy. Sectioning takes a small slice of an otherwise large specimen, and then subsequent antibody staining can be performed. Typically, this is preferable when one desires to stain certain cell types/layers or the entire specimen is too thick (large Z plane) to mount on a coverslip and resolve by microscopy.
Example Question #33 : Gre Subject Test: Biochemistry, Cell, And Molecular Biology
A student researcher runs an agarose gel electrophoresis of a restriction enzyme digested plasmid to generate two fragments of 1200 bp and 3500 bp. The student then stains the gel with ethidium bromide. Which of the following is true about the bands that are seen on the gel?
The 1200 bp fragment will image more brightly than the 3500 bp fragment
The 1200 bp fragment will not image as brightly as the 3500 bp fragment
A simple gel electrophoresis is not sufficient to separate a 1200 bp fragment from a 3500 bp fragment
The 1200 bp fragment will be closer to the loading wells than the 3500 bp fragment when the gel is imaged
None of the other answers
The 1200 bp fragment will not image as brightly as the 3500 bp fragment
The correct answer is the 1200 bp fragment will not image as brightly as the 3500 bp fragment. Ethidium bromide intercalates into DNA proportionally to the size of the fragment. Since we expect the same number of molecules of 1200 bp and 3500 bp fragments, the 1200 bp fragment should be roughly 3 times less bright than the 3500 bp fragment based on the number of ethidium bromide molecules incorporated into the strands. Additionally, smaller fragments migrate more quickly than larger fragments, and as such, are farther from the loading wells.
Example Question #4 : Help With Identification Techniques
When screening a microenvironment for its bacterial species composition, which genes are most commonly used during molecular typing?
Transcription factor genes
DNA polymerase genes
Mitochondrial genes
Transfer RNA genes
Ribosomal RNA genes
Ribosomal RNA genes
The correct answer is ribosomal RNA genes. These genes have highly conserved regions that scientists can use to design primers to amplify intervening regions. These intervening regions, however, are highly divergent and are extremely useful sequences to distinguish between species.
Example Question #31 : Gre Subject Test: Biochemistry, Cell, And Molecular Biology
The minimum distance between two points that can still be seen as distinct structures is termed __________.
contrast
wavelength
resolution
fluorescence
enhancement
resolution
The resolution of an image is defined as the minimum distance between two points so that they can still be seen as two distinct structures. Resolution is important in microscopy. Images with clear resolution can be distinctly seen as two separate objects within the field of view.
Example Question #1 : Diagrams, Graphs, And Tables
A researcher performs a Bradford assay to determine the quantity of an unknown protein in his sample. The standard protein returns absorbance values of 0.101, 0.204, 0.302, 0.405 for the respective quantities of 10ug, 20ug, 30ug, and 40ug of protein. The unknown sample returns an absorbance value of 0.265. What is the quantity of protein in the unknown sample?
For this problem we need to determine the equation of our standard curve. This can be done by creating a graph from the data points and determining the slope.
Assuming that a sample of zero concentration will also have zero absorbance, we can find the equation for the line generated by finding the slope.
Pick two points on the line to find the slope. We will use (0,0) and (40,0.405).
Use this equation and the absorbance given in the question to find the concentration of the unknown sample.
Example Question #1 : Help With Quantification Techniques
Which of the following techniques would help quantify the amount of protein in a sample?
Southern blot
Coimmunoprecipitation
qPCR
Bradford assay
Bradford assay
Bradford assays use shifts in absorption ranges to identify the quantity of a protein in a sample. The technique uses Coomassie blue dye (Bradford reagent) and spectrophotometry to analyze protein concentrations.
qPCR is a procedure that measures the products made from a PCR reaction in real time, and is a helpful tool for studying gene expression. Coimmunoprecipitation is a technique that will isolate a specific protein or protein complex by using an antibody specific to that protein; however, this technique will not quantify a sample without further analysis. Southern blots are used to detect specific DNA sequences in a sample.
Example Question #2 : Help With Quantification Techniques
A researcher wants to know the exact concentration of protein in his sample. He estimates the amount of protein to be between and . Which of the following sets of known protein amounts should he use to set up a standard curve for a Bradford assay?
Because he is unaware of exactly where the concentration lies, it would be best to test a few samples before and after his predicted range. This will allow the researcher to ensure that his curve is linear and that his sample fits accurately somewhere along the curve. If his sample falls outside of the range he tested, he will not be able to use his standard curve to accurately predict the concentration of his sample.
Example Question #2 : Help With Quantification Techniques
A researcher wants to measure the purity of DNA using a spectrophotometer. What wavelength(s) should he/she consider?
A260
The ratio of A280/A260
A280
The ratio of A160/A780
The ratio of A260/A280
The ratio of A260/A280
The correct answer is the ratio of A260/A280. DNA absorbs light most strongly at 260nm and is used to determine the concentration of the DNA solution. Contaminants such as aromatic amino acids present in proteins are absorbed at 280nm, and as such, taking the ratio of A260/A280 will indicate the purity of the DNA sample. Typically, a A260/A280 ratio of 1.7-2 is indicative of good quality DNA.
Example Question #3 : Help With Quantification Techniques
What is the purpose of a Bradford Assay?
To quantify RNA concentration in a sample
To determine the percentage of viable cell culture cells in a population
To quantify DNA concentration in a sample
To quantify protein concentration in a sample
To determine the percentage of cells undergoing apoptosis
To quantify protein concentration in a sample
The correct answer is to determine the concentration of protein in a sample. Bradford assays are often used before Western blots in order to normalize the amount of protein being loaded between conditions. Briefy, a small volume of protein lysate is mixed with a reagent and the absorbance at 595nm is compared to known concentration controls to determine the relative protein concentration of the sample.