11 Questions
Which of the following techniques is commonly used to separate proteins from complex mixtures during purification?
Chromatography
After a production run, a band appears at the expected molecular weight on an SDS-PAGE gel. Which technique is the best way to verify that the band is the purified protein of interest?
Run a Western blot with a sample of the fraction
Which of the following is commonly used in diagnostic immunoassays to quickly diagnose infectious diseases?
Antigens from the pathogen
Which type of ELISA is the most appropriate to determine the concentration of a specific antigen in a sample?
Quantitative ELISA (how much is there)
Which of the following strategies will give you the best resolution of the PCR products on an agarose gel?
Use a higher percentage of agarose and a lower running voltage
In agarose gel electrophoresis, where is the negative electrode terminal located?
At the bottom of the gel (furthest from the well)
If the DNA in lane 3 was loaded on the gel after it was digested with EcoRI, how many EcoRI sites are in the plasmid if the digestion was complete?
2
Which restriction enzyme should the technician use to digest the DNA for cloning the gene of interest into the plasmid?
EcoRI
Which of the following techniques destroys all microorganisms?
Autoclaving
Which focus knob should you use when using a microscope on the highest power objective?
Fine
A solution with a pH of 3.0 is how many times more acidic than a solution with a pH of 6.0?
1000
Study Notes
Protein Purification
- Chromatography is commonly used to separate proteins from complex mixtures during purification.
SDS-PAGE Verification
- Verifying the purified protein of interest after an SDS-PAGE gel run is best done by Western blotting or Mass Spectrometry.
Diagnostic Immunoassays
- ELISA (Enzyme-Linked Immunosorbent Assay) is commonly used in diagnostic immunoassays to quickly diagnose infectious diseases.
ELISA Types
- Sandwich ELISA is the most appropriate type to determine the concentration of a specific antigen in a sample.
PCR Product Resolution
- Using a higher concentration of agarose in the gel gives the best resolution of PCR products on an agarose gel.
Agarose Gel Electrophoresis
- The negative electrode terminal is located at the black electrode (cathode) in agarose gel electrophoresis.
Restriction Enzyme Digestion
- If the DNA in lane 3 was loaded on the gel after being digested with EcoRI and shows multiple bands, it indicates that there are multiple EcoRI sites in the plasmid.
- The number of EcoRI sites can be determined by counting the number of bands.
Cloning and Restriction Enzymes
- The technician should use the same restriction enzyme (EcoRI) to digest the DNA for cloning the gene of interest into the plasmid.
Sterilization Techniques
- Autoclaving is a technique that destroys all microorganisms.
Microscopy
- When using a microscope on the highest power objective, the fine focus knob should be used.
pH Levels
- A solution with a pH of 3.0 is 1000 times more acidic than a solution with a pH of 6.0.
Test your knowledge on aseptic techniques in cell culture. Learn why technicians must practice aseptic techniques and how it ensures the growth of pure cultures and the use of correct media, temperature, and supplements. Discover which technique destroys all microorganisms and more!
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