This set of Gene Manipulation online quiz focuses on “Basic Laboratory Techniques – 3”.
1. What is used to transfer nucleic acid from gels to membranes for further analysis?
a) Gel electrophoresis
Explanation: Blotting is used to transfer nucleic acids from gels to membranes for further analysis. Nucleic acid labeling and hybridization have formed the basis for a range of experimental techniques.
2. Blotting describes the __________ of nucleic acids.
Explanation: Blotting describes the immobilization of sample nucleic acids on to a solid support, generally on nylon or nitrocellulose membrane.
3. Which membrane is used in blotting?
Explanation: The main aim of blotting is immobilization of the nucleic acid samples on a membrane. The membrane must promote binding.
4. Which of the following is used as targets in the blotting techniques?
a) Nucleic acids
d) E. coli
Explanation: The blotted nucleic acids are then used as targets in subsequent hybridization experiments. There are a few blotting procedures.
5. Which of the following is used for the analysis of compositional properties of DNA?
a) Southern blotting
b) Northern blotting
Explanation: Southern blotting is the method used to transfer DNA from agarose gels to membranes so that the compositional properties of DNA can be analyzed.
6. When was the original method of southern blotting developed?
Explanation: The original method for southern blotting was developed in 1975, for detecting fragments in an agarose gel that are complementary to a given RNA or DNA.
7. Southern blotting cannot be used for RNA molecules.
Explanation: Southern method was developed as a variation to the Northern method of DNA analysis. Southern blotting is solely used for RNA.
8. In southern blotting ___________ is present in the reservoir.
Explanation: In the southern blotting technique, the agarose gel is mounted on filter paper wick which dips into a reservoir containing a transfer buffer.
9. The DNA molecules are immobilized on the _____________ in southern blotting technique.
Explanation: The DNA molecules are carried out of the gel by the buffer flow and immobilized on the membrane. Initially, nitrocellulose was used as the membrane.
10. Which membranes have greater binding capacity than nitrocellulose membranes?
Explanation: The main drawback of nitrocellulose membranes is their fragile nature. Supporting nylon membranes have a greater binding capacity and high tensile strength.
11. Larger DNA fragments require a ___________ transfer time.
d) Very high
Explanation: Large DNA fragments, greater than 10 kb require a longer transfer time than short fragments. Uniform transfer hence must be allowed.
12. The depurination treatment in blotting, involves the use of HCl and ____________
d) Nucleic acids
Explanation: To allow uniform transfer of a wide range of DNA fragment sizes, the electrophoresed DNA is exposed to short depurination treatment with HCl followed by alkali.
13. The depurination treatment __________ the DNA fragments.
Explanation: The depurination treatment shortens the DNA fragments by alkaline hydrolysis at depurinated sites. It also denatures the fragments.
14. Depurination denatures the DNA fragments.
Explanation: Depurination denatures the fragments prior to transfer, ensuring that they are in the single stranded state and accessible for probing.
15. Gel is _____________ in neutralizing solution prior to blotting.
Explanation: The gel is equilibrated in neutralizing solution prior to blotting. This is the final step in the southern blotting technique.
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