BIOE 206 Exam 1 UPDATED ACTUAL Questions and CORRECT Answers
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Course
BIOE 206
Institution
BIOE 206
BIOE 206 Exam 1 UPDATED ACTUAL
Questions and CORRECT Answers
Plasmid DNA purification steps - CORRECT ANSWER- 1. grow the bacteria with the
plasmid
2. centrifuge
3. remove supernatant
4. resuspend pellet in lysis buffer (detergents and high salt)
5. neutralize the lysis buffer
6. spin do...
BIOE 206 Exam 1 UPDATED ACTUAL
Questions and CORRECT Answers
Plasmid DNA purification steps - CORRECT ANSWER✔✔- 1. grow the bacteria with the
plasmid
2. centrifuge
3. remove supernatant
4. resuspend pellet in lysis buffer (detergents and high salt)
5. neutralize the lysis buffer
6. spin down to remove debris and genomic DNA
7. put the suspension on a spin column
8. plasmid DNA will adhere to spin column and everything else will be washed away with
high salt buffers
9. add low salt buffer to remove plasmid DNA from spin column
RNA purification steps - CORRECT ANSWER✔✔- Same steps as DNA purification, but
also need to heat to disrupt any secondary structures.
Since the mRNA is the only type that has a polyA tail, the beads can have a "polyT tail"
(oligo dT) that will hybridize to the mRNA and keep it in the column.
Then wash to remove everything that is not mRNA.
Wash with high salt buffer to release the mRNA.
https://www.youtube.com/watch?v=yzqhPBAvvY0
Gel Electrophoresis - CORRECT ANSWER✔✔- Used to separate DNA fragments of
different sizes.
,1. prepare gel
2. load DNA
3. run gel
4. visualize
What direction does the DNA move during gel electrophoresis? Why? - CORRECT
ANSWER✔✔- DNA moves toward cathode (+) because it is negatively charged (PO4-
backbone)
Agarose gel - CORRECT ANSWER✔✔- Used for gel electrophoresis because it has small
pores that allow for DNA movement
For large fragments of DNA, do you want a high or low agarose concentration? - CORRECT
ANSWER✔✔- Low. This will allow the large fragments to move farther apart and it will be
easier to visualize them (.7%)
For small fragments of DNA, do you want a high or low agarose concentration? - CORRECT
ANSWER✔✔- High. This will allow for better visualization of the small fragments. (1.5%)
DNA staining - CORRECT ANSWER✔✔- Ethinium bromide is a carcinogen because it
intercalates into the DNA.
Karyotyping - CORRECT ANSWER✔✔- Allows for chromosomal visualization.
What are the bands in karyotyping? - CORRECT ANSWER✔✔- The light bands are GC rich
areas and the dark bands are AT rich areas.
What can karyotyping detect? - CORRECT ANSWER✔✔- Large chromosomal deletions,
insertions, and translocations. Can diagnose diseases like Down Syndrome (extra 21st
chromosome) or Turner's Syndrome (missing an X chromosome)
Microdissection of human chromosome - CORRECT ANSWER✔✔- Stop the cell cycle
during M phase (when DNA is most condense) and then break the nucleus and locate band of
interest
, FISH - CORRECT ANSWER✔✔- Used for detection of chromosomal rearrangements. Can
detect deletion, amplification, translocation
Can FISH only be used with DNA - CORRECT ANSWER✔✔- No - can also be used with
RNA to study gene expression (mRNA)
FISH steps - CORRECT ANSWER✔✔- 1. make probe ssDNA (complementary with DNA of
interest) with fluorescence tags
2. denature probe and target DNA
3. wash target with probes and allow to hybridize
4. visualize
FISH deletion diagnosis - CORRECT ANSWER✔✔- PTEN (tumor supressor) can be deleted
and FISH can detect this. Label the target DNA with probes for the PTEN gene and visualize
whether there are genes in the chromosomes
FISH amplification diagnosis - CORRECT ANSWER✔✔- C-MYC (type of leukemia) can be
diagnosed with FISH.
FISH translocation diagnosis - CORRECT ANSWER✔✔- CML (type of leukemia) can be
diagnosed with FISH. results from fusion of two genes. Mark those two genes and (red and
green) and look for any yellow overlap in target DNA. Gleevec is a blocker for this fusion
DNA sequencing - Sanger Sequencing - CORRECT ANSWER✔✔- 1. break DNA into
smaller fragments
2. anneal primer
3. carry out 4 rxns (one with each halting neucleotide)
4. run gel
5. you can then figure out the sequence of the complimentary strand
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