BIOL 330 Study Guide - Final Guide: C-Terminus, National Center For Biotechnology Information, Exon

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23 Feb 2017

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Pcr- written notes (page 473)- figure 8-36 & written notes. Pcr is a way to amplify a small region of dna: doesn"t work on rna. 1: separate dna strands (denature them, create a set of primers that will flank the region you are interested in, the primers delineate the exact region you are interested in. Cycle 3: take the two newly synthesized strands and add the two primers to them, you have now produced the first double-stranded copies of your targeted region. 2: cloning a gene encoding a known protein. Primers can be designed from sequence of amino acids or gene sequence. Amplified product can be used as a probe to pull out the full length gene from a cdna or genomic library: amplifying "old dna" Amplifying dna sequences from museum material or fossils - look at evolution of gene sequences (molecular evolution studies: amplifying cloned dna from vectors.

Related Questions

1. You are sequencing a 350bp lizard gene using the Sanger dideoxy sequencing method. You have amplified the region using the primer 5'-ATCCGGGCG-3'. What will be the length of the lowest band in your sequencing gel?

2. Which of the following answer(s) are true (select all that apply)?

Question options:

PCR results in exclusively the desired region to be amplified.

PCR primers must be outside the region of interest. PCR primers must be flanking the region of interest.

Sequencing primers must be flanking the region of interest.

All of the nucleotides used in Sanger Sequencing would also be able to be used for a PCR reaction where you are trying to amplify a gene of interest.

Sanger sequencing requires dNTPs.

A standard PCR reaction requires ddNTPs Small DNA fragments on a gel would be closer to the wells on a gel than larger DNA fragments.

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

Question 1. You are sequencing a 350bp lizard gene using the Sanger dideoxy sequencing method. You have amplified the region using the primer 5'-ATCCGGGCG-3'. What will be the length of the lowest band in your sequencing gel?

(A) Cannot answer the question with the provided information

(B) 350

(D) 9

(E) 351

Question 2. True or False

If cell has a mutated gyrase, it means that it will be unable to properly join together okazaki fragments and therefore will result in many fragmented pieces of DNA.

Question 3. Which of the following answer(s) are true (select all that apply)?

- PCR results in exclusively the desired region to be amplified.

- PCR primers must be outside the region of interest.

- PCR primers must be flanking the region of interest.

- Sequencing primers must be flanking the region of interest.

- All of the nucleotides used in Sanger Sequencing would also be able to be used for a PCR reaction where you are trying to amplify a gene of interest.

- Sanger sequencing requires dNTPs.

- A standard PCR reaction requires ddNTPs

- Small DNA fragments on a gel would be closer to the wells on a gel than larger DNA fragments.

BIOL 330 Study Guide - Final Guide: C-Terminus, National Center For Biotechnology Information, Exon

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