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1) In general, the length of the primer is 18 -25 nucleotides. Why? 2) Thermostable DNA polymerase commonly used in PCR is Taq polymerase and Pfu polymerase. Compare these characteristics.

please explain in details!!! thank you

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Irving Heathcote
Irving HeathcoteLv2
29 Sep 2019

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Related questions

The PCR (polymerase chain reaction) protocol that is currently used in laboratories was facilitated by the discovery of a bacterium called Thermus aquaticus in a hot spring inside Yellowstone National Park, in Wyoming. This organism contains a heat-stable form of DNA polymerase known as Taq polymerase, which continues to function even after it has been heated to 95°C. Why would such a heat-stable polymerase be beneficial in PCR?

What would happen if it weren’t heat stable?

How might you choose a region of DNA for a PCR primer so as to increase the temperature necessary for primer annealing (to minimize nonspecific PCR products)?

A PCR reaction begins with 5 double stranded segment of DNA. Estimate the number of double-stranded copies of DNA that are present after the completion of 15 amplification cycles?

Question 1 (3 points)

Explain why you need two primers for PCR and what would happen if you did not include 2 in your reaction.

What component(s) of your PCR reaction give it specifity? In other words, what reagent will you add to your PCR reaction that will allow it to amplify a specific piece of DNA and not all DNA in general?

a. Buffer

b. dNTPs

c. Primers

d. Taq Polymerase

If you failed to input the proper annealing temperature on the thermocycler, what is the most likely outcome?

a. Taq polymerase will not actively polymerize DNA so no PCR product will be visible on a gel.

b. DNA will not denature properly so PCR will work, just not as efficiently as it should.

c. Primers will not bind to the denatured DNA so little or no PCR product will be visible on a gel.

d. DNA will be degraded so no PCR product will be visble on a gel.

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 94°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 55°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 72°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

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