D 12) Which of these correctly explains how DNA migrates through an agarose gel? a. Longer...

  

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D 12) Which of these correctly explains how DNA migrates through an agarose gel? a. Longer fragments migrate faster because they have more charges to be repelled from one pole and attracted to the other b. The relative migration rate is determined primarily by the GC content, with high GC content tragments travelling more slowly c. The relative migration rate is determined primarily by the isoelectric point, which depends on erice and must be empirically determined the d. Short ent migrate faster because they have less resistance traveling through the matrix/pres of the gel 13) Sanger DNA sequencing is also called chain termination sequencing. Why is this? า part 2 a. Modified nucleotides lacking a 3' OH are added to the reaction b. c. A low processivity polymerase is used that tends to fall of the template before about 1000bp A limiting quantity of dNTPs are added, such that each DNA polymerase will not be able to complete the full-length synthesis d. A short time is allowed for elongation, such that the DNA polymerases will be unable to complete full-length synthesis 14) In modern Sanger DNA sequencing, how is the sequence determined following the PCR? a. Mass spectrometry; PCR products are randomly fragmented and the masses of the overlapping fragments are compared to identify the order of the nucleotides b. Probe hybridization; PCR products are allowed to hybridize to immobilized probes of known, overlapping sequences, by comparing the probes that interact with the product one can compute the most probably sequence of nucleotides Fluorescence detection; the last-incorporated nucleotide is fluorescent with a color unique to the type of base it has d. Fluorescence detection; each nitrogenous base has a slightly different intrinsic fluorescence, and by comparing the fluorescence readings of the different products once can determine if, for instance, one product had one more adenine than another, and that must be the last incorporated nucleotide 15) You PCR amplify a typical protein-coding gene from human genomic DNA and ligate the sequence into an E. coli expression plasmid. What do you anticipate? a. The E. coli cell will express the protein with the same amino acid sequence as in humans because the code is interpreted identically b. Since genomic DNA was used as template, the presence of introns is likely and the correct, full- length protein would not be expected c. The E. coli cell will express the protein with the correct amino acid sequence, but post- translational modifications unique to eukaryotes will be missing d. No mRNA or protein product will be made, because a eukaryotic protein coding sequence will be unable to be transcribed by prokaryotic RNA polymerase D 12) Which of these correctly explains how DNA migrates through an agarose gel? a. Longer fragments migrate faster because they have more charges to be repelled from one pole and attracted to the other b. The relative migration rate is determined primarily by the GC content, with high GC content tragments travelling more slowly c. The relative migration rate is determined primarily by the isoelectric point, which depends on erice and must be empirically determined the d. Short ent migrate faster because they have less resistance traveling through the matrix/pres of the gel 13) Sanger DNA sequencing is also called chain termination sequencing. Why is this? า part 2 a. Modified nucleotides lacking a 3' OH are added to the reaction b. c. A low processivity polymerase is used that tends to fall of the template before about 1000bp A limiting quantity of dNTPs are added, such that each DNA polymerase will not be able to complete the full-length synthesis d. A short time is allowed for elongation, such that the DNA polymerases will be unable to complete full-length synthesis 14) In modern Sanger DNA sequencing, how is the sequence determined following the PCR? a. Mass spectrometry; PCR products are randomly fragmented and the masses of the overlapping fragments are compared to identify the order of the nucleotides b. Probe hybridization; PCR products are allowed to hybridize to immobilized probes of known, overlapping sequences, by comparing the probes that interact with the product one can compute the most probably sequence of nucleotides Fluorescence detection; the last-incorporated nucleotide is fluorescent with a color unique to the type of base it has d. Fluorescence detection; each nitrogenous base has a slightly different intrinsic fluorescence, and by comparing the fluorescence readings of the different products once can determine if, for instance, one product had one more adenine than another, and that must be the last incorporated nucleotide 15) You PCR amplify a typical protein-coding gene from human genomic DNA and ligate the sequence into an E. coli expression plasmid. What do you anticipate? a. The E. coli cell will express the protein with the same amino acid sequence as in humans because the code is interpreted identically b. Since genomic DNA was used as template, the presence of introns is likely and the correct, full- length protein would not be expected c. The E. coli cell will express the protein with the correct amino acid sequence, but post- translational modifications unique to eukaryotes will be missing d. No mRNA or protein product will be made, because a eukaryotic protein coding sequence will be unable to be transcribed by prokaryotic RNA polymerase

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