CGMBS Analysis and Characterization of Nucleic Acids and Proteins Lecture Questions And Answers Rated 100% Correct!!
0 view 0 purchase
Course
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
The benefits of buying summaries with Stuvia:
Guaranteed quality through customer reviews
Stuvia customers have reviewed more than 700,000 summaries. This how you know that you are buying the best documents.
Quick and easy check-out
You can quickly pay through credit card or Stuvia-credit for the summaries. There is no membership needed.
Focus on what matters
Your fellow students write the study notes themselves, which is why the documents are always reliable and up-to-date. This ensures you quickly get to the core!
Frequently asked questions
What do I get when I buy this document?
You get a PDF, available immediately after your purchase. The purchased document is accessible anytime, anywhere and indefinitely through your profile.
Satisfaction guarantee: how does it work?
Our satisfaction guarantee ensures that you always find a study document that suits you well. You fill out a form, and our customer service team takes care of the rest.
Who am I buying these notes from?
Stuvia is a marketplace, so you are not buying this document from us, but from seller PatrickKaylian. Stuvia facilitates payment to the seller.
Will I be stuck with a subscription?
No, you only buy these notes for $7.99. You're not tied to anything after your purchase.
