BIOL 1002 : Test 5

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azuremammoth471

15 Mar 2019

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The probability of finding a recognition site can be calculated if the length of the recognition site is known. How many times would you expect to find a 6 base pair recognition site in a dna fragment of. In an experiment, an ampicillin-resistant plasmid vector was used to introduce a piece of foreign. The key difference between a retrovirus and other viruses is that retroviruses are. Culture the cells in a medium containing ampicillin. Rna-based, and use reverse transcriptase to form dna which can integrate into the host genome. While the dna sequence necessary for the production of a particular protein in prokaryotes can be determined directly from the chromosomal dna, this is not possible in eukaryotes. If a gene from a eukaryotic cell were cloned directly from its chromosomal. Which of the following correctly matches the macromolecule with its length in nucleotides or amino acids? cdna is.

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1) Which of the following is FALSE about plasmid vectors ued in recombinant DNA research?
A) Usually carrry an antibiotic resistance gene
B) Clonin sites are always located between genes, that is, outside coding sequences
C) May hae a multiple clonin gsite which can be cut by several restriction enzymes
D) May be expression vectors

2) Which of the following is FLASE about Sanger Dideoxy DNA sequencing protocols?
A) Different specfiic chemical reactions break the DNA at each base.
B) Fluorescent molecules can be used to detect the DNA
C) The Template is single stranded DNA
D) Many steps can be automated

3) A southern blot transfers ___ to a Nitrocellulose membrane.
A) Protein
B) RNA
C) DNA
D) Lipid

4) Which of the following is true about cDNA?
A) It is made from DNA template
B) It is isolated from bacteria
C) Reverse transcriptase is used to synthesize cDNA
D) It has introns

5) The restriction endonucleases used in recombinant DNA work:
A) Always produce blunt ends (No unpaired bases)
B) Recognize sequences 4-6 bp long
C) Cut the DNA outisde the recognition sequence
D) All of the above

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1. You have cloned a human gene along with its promoter and ribosome-binding site in a bacterial cloning vector and then introduced the recombinant plasmid into a bacterial host cell that supports the replication and segregation of the cloning vector. Unfortunately, even though you know the whole human gene sequence is present (by DNA hybridization); you cannot get expression of the gene. Which of the following is NOT a reason why there is no expression?

A. The vector cannot replicate

B. The human promoter is not recognized by the bacterial host cell

C. The ribosome-binding site is not recognized by the bacterial host cell

D. The human gene lacks introns, so it is not transcribed by the host bacterium

2. Restriction endonucleases are:

A. sequence-specific

B. able to recognize any 4-, 6-, or 8-base pair sequence

C. always able to produce single stranded tails

D. only able to cleave bacterial DNA

3. Which of the following is NOT a required characteristic of cloning vectors?

A. Genes for replication of vector DNA and segregation to progeny cells

B. Genes for selection of cells carrying the vector (e.g., antibiotic-resistance)

C. DNA sequence for the introduction of foreign DNA where the integrated DNA does not disable the vector

D. None of the above

4. A poly (T) primer is used to produce cDNA from mammalian mRNA. Which of the following would be used to produce cDNA from bacterial mRNA?

A. A poly (T) primer

B. A random DNA sequence primer

C. A poly (A) primer

D. A poly (GC) primer

1.) You measure the OD600 of your culture of chloramphenicol-treated XL1-Blue/pTrc99A cells as described in Step 1 of Experiment 3A and obtain a value of 0.265. Assuming a plasmid copy number of 570 plasmids per cell, and a 100% recovery of plasmid DNA from 3.00 mL of the cells, what is the theoretical yield of plasmid DNA in micrograms (ug)? 2 points (Note: the size of pTrc99A is 4176 base pairs and the average molecular weight of double-stranded DNA is 660 g/mole of base pairs; report your answer with three significant figures) Tries 0/3

2.) Would you expect your actual yield of DNA to be more or less than the theoretical yield? 1 point Tries 0/1

3.) Assuming 100% recovery of DNA from the phage DNA isolation (Experiment 3B) and considering that not all the phage in your stock are able to infect E. coli successfully, would you expect your actual yield of DNA to be more or less than the theoretical yield calculated from your titer of the phage stock? 1 point Tries 0/1

4. Choose (True or False) whether each of the following parameters effects the rate of migration of DNA through agarose gels. 2 points Length of time in electric field DNA shape (supercoiled vs. relaxed circular vs. linear)

Volume of sample loaded (assuming all of the sample is contained within the loading well and enters the gel)

Electric field strength (volts per centimeter)

DNA size (molecular weight)

Physical length of the agarose gel

Polarity of the electric field Porosity of gel (% (w/v) agarose) Tries 0/3

5.) Ethidium bromide stains DNA by intercalating between adjacent base pairs, resulting in an increase in fluorescence, and can be used to quantify the mass of DNA present in a band of DNA on an agarose gel. Will the staining intensity of 100 ng of supercoiled plasmid DNA be the same as 100 ng of the plasmid that has been linearized by a single-site digestion with a restriction endonuclease?1 point Tries 0/1

6.) Choose (True or False) whether each of the following parameters effects the distance of migration of DNA through agarose gels. 1 point Polarity of the electric field

DNA size (molecular weight)

Length of time in electric field

Electric field strength (volts per centimeter)

Volume of sample loaded (assuming all of the sample is contained within the loading well and enters the gel)

Physical length of the agarose gel

Porosity of gel (% (w/v) agarose)

DNA shape (supercoiled vs. relaxed circular vs. linear)

BIOL 1002 : Test 5

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