BIO SCI 97 Lecture Notes - Lecture 3: Central Dogma Of Molecular Biology, Transfer Rna, Ibm 7090

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30 Jan 2014

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Pcr: requires a dna template, requires a sequence-specific primer you have to know at least part of the dna sequence, requires nucleotide triphosphates, requires a special polymerase. Dna separates into two strands of dna by heating to 95c. G-c has 3 h-bonds and a-t have 2-h bonds so diff. organisms have different heating points. After separating, you cool the dna and the short primers that are between 16-25 bases long and they base pair to the two single strands. Each of the strands will base pair with a primer. Raise the temperature slightly so the dna polymerase will start off at the primer and copy the two strands. 95 c denatures dna by breaking the hydrogen bonds between strands. 50-65 c temperature is reduced to allow primer to bind (anneal) to dna. 72 c temperature is raised to the optimum for the thermostable polymerase (taq) to extend the dna strand.

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

Double check to see if I got these right? 3 questions

Some Processes Involved in DNA Replication

1.The replication origin is identified.

2 DNA primase builds RNA primer.

3Okazaki fragments are spliced by DNA ligase.

4Helicases bind and uncoil the DNA double helix.

5 RNA primer is the starting point for DNA polymerase.

6 DNA replication ends at the telomere where DNA codes fortermination.

7 DNA polymerase adds complementary nucleotides in the 5' to 3'direction in short segments called Okazaki fragments.

The sequence in which the events numbered above occur during DNAreplication of the lagging strand: Number 1 - 7 in order.

I put: 1,4,2,5,7,3,6

__________________Question 2:

Some Functions of Enzymes

Build DNA strands
Build RNA primers
Build mRNA strand
Cleave DNA strands
Splice Okazaki fragments

Enzyme:
Helicase :4
DNA ligase :5
RNA primase :1
RNA polymerase :3
DNA polymerase:2

Question 3:

like mRNA, tRNA has a ribose sugar, U instead of T, and issingle stranded. Unlike mRNA, which remains a long single strand ofnucleotides, tRNA folds so that some areas pair up.

The resulting structure has an anticodon on one end and a site foran amino acid to attach on the other end. There is basecomplementarity (A pairs with U and G pairs with C) between an mRNAcodon and tRNA anticodon.

If the amino acid tyrosine attaches to a tRNA, which of thefollowing anticodons could be at the opposite end of the tRNAmolecule?

Select one:

a. ATA and ATG

b. UAU and UAC

c. AUA and AUG

d. AGA and AGU

I picked B.

purpleturtle13

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

BIO SCI 97 Lecture Notes - Lecture 3: Central Dogma Of Molecular Biology, Transfer Rna, Ibm 7090

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