BIOLOGY 1A03 Lecture : Biology Chapter 19.docx

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The polymerase chain reaction (pcr) is an in vitro dna synthesis reaction in which a specific section of dna is replicated over and over to amplify the number of copies of that sequence. It"s a technique for generating many identical copies of a particular section of dna. Although pcr is much faster and technologically easier than cloning genes into a dna library, it is only possible when a researcher has some information about dna sequences near the gene in question. Sequence information is required because to do a polymerase chain reaction, you have to start with short lengths of a single-stranded dna that match sequences on either side of the gene of interest. These short segments act as primers for the synthesis reaction. Primer sequences must be complementary to bases on either side of the gene in question. One primer is complementary to a sequence on one strand upstream of the target dna, the sequence of interest.

Related Questions

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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

PCR Amplification, sequencing and Genetic identification of Prokaryotes

1. Compare DNA replication in a typical cell to the synthetic DNA replication that happens during the Polymerase Chain Reaction (PCR). Fill in the missing information that describes how each process is accomplished (i.e. enzyme name or other method) in each. Two answers are given as example:

Event	Cell	PCR
Unwinding of DNA Strands	DNA Helicase
Primer Synthesis		Synthetic primers added
New DNA Strand synthesis

2. Heating proteins to 94Â°C would destroy the function of most human or bacterial enzymes, including DNA Polymerase. How is this avoided during the DNA strand synthesis step in PCR?

3. What are the two major factors that influence the rate at which DNA amplicons move through an electrophoresis gel?

4. What are the three most significant features of the 16S rRNA gene that make it an excellent choice for DNA homology analysis and identification in bacteria?

aquamarinecamel49

1)What enzyme is used to attach different colors of stained nucleotides to create a DNA molecule along the mRNA?

Select one:

a. reverse transcriptase

b. fluorescent primer

c. restriction enzyme

d. radioactive probe

The process in which two complementary strands of DNA from different sources pair with one another is called hybridization.

Select one:

True

False

3) Explain the significance of a red microarray spot revealed by the microarray scanner.

Select one:

a. this gene is not expressed in either type of cell

b. this gene is expressed in cancer cells

c. this gene is expressed in healthy cells and cancer cells

d. this gene is expressed in healthy cells

4)What is the function of primers in PCR?

Select one:

a. raw material for new DNA

b. attach new nucleotides to DNA strand

c. attach to either side of target DNA

d. denature, or separate, DNA

5)What is the function of DNA polymerase in PCR?

Select one:

a. raw material for new DNA

b. attaches to either side of target DNA

c. attaches new nucleotides to DNA strand

d. denatures, or separates, DNA

BIOLOGY 1A03 Lecture : Biology Chapter 19.docx

BIOLOGY 1A03 Full Course Notes

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