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BIO 206 exam 1 Practice Questions and Answers (100% Pass)
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BIO 206 exam 1 Practice Questions and Answers (100% Pass)
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©PREP4EXAMS@2024 [REAL EXAM DUMPS] Tuesday, July 23, 2024 10:52 AMBIO 206 exam 1 Practice Questions and Answers (100% Pass)
Plasmid DNA purification steps - ✔️✔️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
https://www.jove.com/science-education/5062/plasmid-purification
RNA purification steps - ✔️✔️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 - ✔️✔️Used to separate DNA fragments of different sizes.
1
, ©PREP4EXAMS@2024 [REAL EXAM DUMPS] Tuesday, July 23, 2024 10:52 AM
1. prepare gel
2. load DNA
3. run gel
4. visualize
What direction does the DNA move during gel electrophoresis? Why? - ✔️✔️DNA moves toward
cathode (+) because it is negatively charged (PO4- backbone)
Agarose gel - ✔️✔️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? - ✔️✔️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? - ✔️✔️High. This
will allow for better visualization of the small fragments. (1.5%)
DNA staining - ✔️✔️Ethinium bromide is a carcinogen because it intercalates into the DNA.
Karyotyping - ✔️✔️Allows for chromosomal visualization.
What are the bands in karyotyping? - ✔️✔️The light bands are GC rich areas and the dark bands
are AT rich areas.
What can karyotyping detect? - ✔️✔️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 - ✔️✔️Stop the cell cycle during M phase (when DNA is
most condense) and then break the nucleus and locate band of interest
FISH - ✔️✔️Used for detection of chromosomal rearrangements. Can detect deletion,
amplification, translocation
Can FISH only be used with DNA - ✔️✔️No - can also be used with RNA to study gene
expression (mRNA)
FISH steps - ✔️✔️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
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