01:119:115 Lecture Notes - Lecture 17: Antimicrobial Resistance, Polyacrylamide Gel Electrophoresis, Chain Termination

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olivekangaroo435

13 Mar 2016

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Base pairing of one strand of nucleic acids to a complementary strand. Genetic engineering= manipulation of genes for practical purposes: dna sequencing. Used to sequence a dna fragment up to about 1000 bp. What do you need to run this method: denature your fragment of dna to be sequenced, primers- short segments w/ known 3" ends (prepare for process, 4 deoxyribonucleotides, 4 dideoxyribonucleotides- labeled w/ florescent molecules (ddntps) New strand sequence starts at 3" end of primer. B/c there is no free oh, the strand cannot elongate. Generates a set of labeled strands of diff. lengths- labeled by color. Polyacrylamide gel- acts as a molecular sieve. Why: explore evolutionary relationships, discover the sequence of dna fragment. Produces by bacteria: natural defense against phages, cut up foreign dna. Recognize & cut at a restriction site: specific short dna sequence, enzymes cut at both ends of site.

Related Questions

31. Why does incorporation of a dideoxynucleotide into a growingDNA polymer terminate strand synthesis?

a. There is no longer a free 5âphosphate to which to add the next nucleotide.

b. There is no longer a free 3â hydroxyl to which to add thenext nucleotide.

c. There is no longer a free 3â phosphate to which to add thenext nucleotide.

d. There is no longer a free 5â hydroxyl to which to add thenext nucleotide.

2. The output from an automatedsequencing reaction shows a series of fluorescent peaks. Whenconsidering the data output, why are some peaks larger thanothers?

a. Chain termination occurred more frequently at the basecorresponding to a large peak, producing a relatively largerfluorescent signal.

b. The primer had multiple bases fluorescently labeled forproducts corresponding to the larger peaks.

c. More bases were fluorescently labeled in the DNA strandscorresponding to the larger peaks.

d. Some fluorescent colors are brighter than others.

3. What allows chain terminationto occur at each base of the template during an automatedsequencing reaction?

a. the presence of a relatively low concentration of ddNTPs

b. the presence of a relatively low concentration of dNTPs

c. the presence of a relatively high concentration of ddNTPs

d. the presence of a relatively high concentration of dNTPs

4. In automated fluorescent sequencing, what would happen if thefluorescently labeled ddATP was inadvertently omitted from thereaction?

a. DNA polymerase does not function without equal concentrationsof the four nucleotides

b there would be gaps in the sequence where thymines werepresent in the template DNA strand

c. the sequence would end where the first thymine wasencountered on the template DNA strand

d. the sequence would end where the first adenine should beincorporated

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01:119:115 Lecture Notes - Lecture 17: Antimicrobial Resistance, Polyacrylamide Gel Electrophoresis, Chain Termination

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