01:119:115 Lecture Notes - Lecture 17: Polyacrylamide Gel Electrophoresis, Molecular Sieve, Restriction Enzyme

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24 Dec 2015

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Dna technology- techniques that involve sequencing or manipulating dna. Dideoxyribose (missing 2 hydroxyl groups: 2" and 3") Each will be labeled with a different fluorescent label: machine primer the strand, synthesis of each new strand to begin at the 3" end of the continues until a dideoxyribonucleotide is inserted. When it is inserted it will prevent the further elongation of: set of labeled strands because there is no free 3" hydroxyl. Restriction enzymes= restriction endonucleases: produced by bacteria. * * * * a 5" and cytosines within the restriction sites adenines: hind iii- enzyme is going to cut. 5" a-a-g-c-t-t 3" =palindrome(read the same both ways) Dna cloning [fig 20. 5: methods for preparing well-defined dna segments in multiple identical copies, cloning methods use escherichia coli (e coli, large circular chromosomes, plasmids smaller, circular dna molecules. Small number of genes not required for survival.

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: Polyacrylamide Gel Electrophoresis, Molecular Sieve, Restriction Enzyme

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