CGMBS Analysis and Characterization of Nucleic Acids and Proteins Lecture Questions And Answers Rated 100% Correct!!
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CGMBS Analysis and Characterization of Nucleic
Institution
CGMBS Analysis And Characterization Of Nucleic
What are the steps of a southern blot? - ️️1. DNA is isolated and cut with restriction
enzymes
2. Fragments are separated by gel electrophoresis
3. DNA fragments are exposed to a labeled probe (complementary DNA or RNA),
complementary to region of interest
What is RFLP? - ️️- Restricti...
CGMBS Analysis and Characterization of
Nucleic Acids and Proteins Lecture
What are the steps of a southern blot? - ✔️✔️1. DNA is isolated and cut with restriction
enzymes
2. Fragments are separated by gel electrophoresis
3. DNA fragments are exposed to a labeled probe (complementary DNA or RNA),
complementary to region of interest
What is RFLP? - ✔️✔️- Restriction Fragment Length Polymorphism
- The number and location of restriction sites for a given restriction enzymes are not the
same in all individuals
What is the mapping difference between linear DNA fragments and circular DNA
fragments? - ✔️✔️Linear makes one more fragment than circular fragments when cut
the same amount
What is CRISPR? - ✔️✔️- Clustered Regularly Interspaced Short Palindromic Repeats
- Classes of repeated DNA sequences matching the genome found in prokaryotic and
archaebacterial genomes
What is CRISPR used for? - ✔️✔️gene editing
How does CRISPR manipulate DNA? - ✔️✔️CRISPR/Cas9 edits genes by precisely
cutting DNA and then letting natural DNA repair processes to take over
What are we looking for in a southern blot? - ✔️✔️Target DNA
What is the purpose of the probe in a southern blot? - ✔️✔️Signal of the probe is
detected to indicate the presence or absence of the sequence
What does properly restricted genomic DNA look like on a gel after electrophoresis? -
✔️✔️Genomic DNA cut with restriction enzymes should produce a smear representing
the billions of fragments of all sizes released by digestion by the enzyme
What does improperly cutting with restriction enzyme result in? - ✔️✔️Incorrect bands
being shown
What does a large aggregate of DNA near the top of the lane indicate? - ✔️✔️indicates
that the restriction enzyme activity was incomplete
, What does a smear located in the lower region of the lane indicate? - ✔️✔️sign that
DNA is degraded
What is depurination? - ✔️✔️- Large fragments need the gel to be soaked in dilute HCl
solution
- This process remove purine bases from the sugar phosphate backbone
- This loosens the larger fragments for more complete denaturation
What is a restriction map? - ✔️✔️a map of known restriction sites within a sequence of
DNA
Why is the denaturation step important in southern blot? - ✔️✔️Single stranded DNA
binds more tightly to nitrocellulose membrane on transfer
What is the purpose of membrane in southern blot? - ✔️✔️The fragments are
permanently attached to the membrane, allowing probes to be added and removed and
new probes to be added
What is capillary transfer? - ✔️✔️movement of buffer from a soaked paper to the dry
paper, denatured DNA moves from the gel to the membrane where the gel is in
between the papers
What is electrophoretic transfer? - ✔️✔️Electric current move DNA from gel to the
membrane. Mostly for small fragment or proteins
What is vacuum transfer? - ✔️✔️System uses suction and buffer recirculation to move
DNA out of the gel to the membrane. Its faster than capillary for larger DNA fragments
What is prehybridization in southern blot? - ✔️✔️to prevent the probe from binding to
nonspecific sites on the membrane surface which can cause a background noise
What are the steps in northern blot? - ✔️✔️1. Gel electrophoresis
2. Transfer (blot)
3. Immobilize
4. Block
5. Probe
6. Wash (2x)
7. Visualize
What are we looking for in northern blot? - ✔️✔️look from specific RNAs using DNA
probes
What are the steps in western blot? - ✔️✔️1. Sample prep
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