BIOLOGY 1A03 Lecture Notes - Lecture 21: Frederick Sanger, Sanger Sequencing, Polymerase Chain Reaction

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BIOLOGY 1A03 Full Course Notes

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Theme 4, module 3: applications of dna replication. Polymerase chain reaction - ability to amplify dna in a tube in 25-35 cycles. Denaturation: high temperatures heat solution causing double helix to unwind. Annealing: solution is cooled so primers can anneal to complementary strands each on an opposite end (5" end of primer attaches to 3" dna end) Extension: the addition of nucleotides; dna grows exponentially. Pcr does not always work (something can go wrong) thus, they compare the size of produced. Dna to the expected size of the dna) *smaller & more negatively charged atoms travel faster: dna fragments dyed and fluoresce when exposed to uv light. Large scale sequencing project that can sequence entire genome known as shotgun sequencing. Being able to break entire genome into different sized pieces then proceeding with 3 specific phases a. b. c. Phase 1: random sequencing of dna in each fragment. Phase 1: dna sequencing *reveals coding and non-coding regions.

Related Questions

Please answer all

Why does a template-dependent DNA polymerase require a primer to initiate DNA synthesis?

	DNA polymerase require 5' phosphate to add a new nucleotide
	DNA polymerase require 3' hydroxyl group to add a new nucleotide

DNA polymerase require energy by hydrolizing the primers

Which of the following enzymes is template independent?

	Reverse transcriptase
	Klenow fragment

	DNA polymerase
	Terminal deoxynucleotidyl transferase

RNA polymerase

The natural function of a DNA ligase enzyme is:

	Joining together DNA sequences with compatible cohesive ends from two different organisms to yield a recombinant DNA molecule
	Sealing single stranded nicks in double stranded DNA molecules

	Joining together two blunt ended fragments of DNA within a cell
	Joining together two single stranded DNA molecules

The natural function of 3' -> 5' exonuclease activity of a DNA polymerase is to:

	Remove the 5' end of the DNA strand that is being copied
	Remove damaged nucleotide from the template strand

	Remove nucleotides from the end of DNA to generate blunt ends
	Remove incorrect nucleotides from the newly synthesized DNA

Which technique is used to resolve the different sizes of DNA fragments?

	Gene cloning
	PCR

	DNA sequencing
	Gel electrophoresis

Which of the following vectors does not contain bacterial origin of replication (oriC)?

	plasmid
	cosmid

	bacterial artificial chromosome
	lambda vector

Which process of bacteriophage is not used for gene cloning?

	Concatemer formation
	Lytic cycle

	Lysogenic cycle
	viral packaging

What vector would be best suited for creating a contig of bovine (cattle) chromosome 10?

	lambda vector
	Cosmid vector

	P1 vector
	Plasmid

E. coli takes up plasmid DNA by which of the following methods?

	Transduction
	Transformation

	Conjugation
	Transfection

The following times and temperatures are an example of the steps for PCR. You can use the Figure to help you answer the following questions.

Why is the first step is carried out at 94Â°C?

	To degrade the template
	To denature the template

	To activate the polymerase
	To facilitate the primer binding

What happens in the reaction when the temperature shifts to 55Â°C during cycling?

	the DNA polymerase will carry out DNA synthesis by extending the annealed primers
	the DNA polymerase will finish DNA synthesis

	the primers anneal to the single-stranded regions of the DNA
	the primers anneal to the double-stranded regions of the DNA

During cycling, what occurs when the temperature is at 72Â°C?

	the primers anneal to the double-stranded regions of the DNA
	the DNA polymerase will carry out DNA synthesis by extending the annealed primers

	the primers anneal to the single-stranded regions of the DNA
	the DNA polymerase will finish DNA synthesis

You designed a set of primers to amplify 1 kb DNA with polymerase chain reaction. By which cycle of polymerase chain reaction, we would expect to see the first double stranded DNA with expected size?

	The end of cycle 1
	The end of cycle 2

	The end of cycle3
	The end of cycle 4

BIOLOGY 1A03 Lecture Notes - Lecture 21: Frederick Sanger, Sanger Sequencing, Polymerase Chain Reaction

BIOLOGY 1A03 Full Course Notes

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