GRE Subject Test: Biochemistry, Cell, and Molecular Biology : Laboratory Practices

Study concepts, example questions & explanations for GRE Subject Test: Biochemistry, Cell, and Molecular Biology

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All GRE Subject Test: Biochemistry, Cell, and Molecular Biology Resources

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Example Questions

Example Question #4 : Help With Identification Techniques

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? 

Possible Answers:

None of the other answers 

Generate the custom antibody against purified protein rather than a predictive peptide

Determine optimal blocking buffer components and concentrations 

Affinity purify the custom antisera

Determine optimal antibody dilution for the application by performing serial dilutions 

Correct answer:

None of the other answers 

Explanation:

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 #5 : Help With Identification Techniques

When is indirect immunofluorescence of sectioned specimens beneficial over whole-mounted specimens? 

Possible Answers:

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

Sectioning is preferred if the whole-mount is too large in the Y plane to resolve by confocal microscopy

Sectioning is preferred if the whole-mount is too large in the X plane to resolve by confocal microscopy

Sectioning if preferred when using RNA probes, whereas whole-mount is solely for antibody staining 

Correct answer:

Sectioning is preferred if the whole-mount is too large in the Z plane to resolve by confocal microscopy

Explanation:

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 #6 : Help With Identification Techniques

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?  

Possible Answers:

A simple gel electrophoresis is not sufficient to separate a 1200 bp fragment from a 3500 bp fragment 

The 1200 bp fragment will image more brightly than the 3500 bp fragment 

None of the other answers 

The 1200 bp fragment will not image as brightly as the 3500 bp fragment

The 1200 bp fragment will be closer to the loading wells than the 3500 bp fragment when the gel is imaged

Correct answer:

The 1200 bp fragment will not image as brightly as the 3500 bp fragment

Explanation:

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 #7 : Help With Identification Techniques

When screening a microenvironment for its bacterial species composition, which genes are most commonly used during molecular typing? 

Possible Answers:

DNA polymerase genes

Ribosomal RNA genes 

Transfer RNA genes 

Transcription factor genes 

Mitochondrial genes

Correct answer:

Ribosomal RNA genes 

Explanation:

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 #11 : Help With Identification Techniques

The minimum distance between two points that can still be seen as distinct structures is termed __________.

Possible Answers:

enhancement

wavelength

fluorescence

contrast

resolution

Correct answer:

resolution

Explanation:

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 : Help With Quantification Techniques

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?

Possible Answers:

Correct answer:

Explanation:

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.

 

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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 #31 : Laboratory Practices

Which of the following techniques would help quantify the amount of protein in a sample?

Possible Answers:

qPCR

Bradford assay

Southern blot

Coimmunoprecipitation

Correct answer:

Bradford assay

Explanation:

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?

Possible Answers:

Correct answer:

Explanation:

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 #32 : Laboratory Practices

A researcher wants to measure the purity of DNA using a spectrophotometer. What wavelength(s) should he/she consider? 

Possible Answers:

A260

The ratio of A160/A780

The ratio of A280/A260

A280

The ratio of A260/A280

Correct answer:

The ratio of A260/A280

Explanation:

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 #33 : Laboratory Practices

What is the purpose of a Bradford Assay? 

Possible Answers:

To determine the percentage of cells undergoing apoptosis

To determine the percentage of viable cell culture cells in a population

To quantify protein concentration in a sample

To quantify DNA concentration in a sample

To quantify RNA concentration in a sample

Correct answer:

To quantify protein concentration in a sample

Explanation:

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. 

All GRE Subject Test: Biochemistry, Cell, and Molecular Biology Resources

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