The use of thermostable DNA (deoxyribonucleic acid) polymerase in PCR (polymerase chain reaction). Introduction: The technique that involves the cloning of the DNA fragment can either use vector (gene cloning) or work without it (PCR). In PCR, the DNA fragments are amplified using the dNTPs (deoxyribonucleotide triphosphate), polymerase, ligases, and primers.

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Definition Definition Chemical process in which one or more monomers combine to produce a very large chain-like molecule called a polymer. The functional groups present on the monomers and their steric effects are responsible for polymerization through a sequence of reactions that vary in complexity. There exists a stable covalent chemical bond between monomers that sets apart polymerization from other processes.

Chapter 21, Problem 9EQ

Summary Introduction

To review:

The use of thermostable DNA (deoxyribonucleic acid) polymerase in PCR (polymerase chain reaction).

Introduction:

The technique that involves the cloning of the DNA fragment can either use vector (gene cloning) or work without it (PCR). In PCR, the DNA fragments are amplified using the dNTPs (deoxyribonucleotide triphosphate), polymerase, ligases, and primers.

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When performing cloning experiments, it is not always necessary to treat sources of DNA with the same restriction enzyme. For example, DNA treated with EcoRI can be combined with DNA from a treatment using FunII. Explain why this is possible.

The plasmid cloning vector pBR322 is cleaved with the restriction endonuclease PstI. An isolated DNA fragment from a eukaryotic genome (also produced by PstI cleavage) is added to the prepared vector and ligated. The mixture of ligated DNAs is then used to transform bacteria, and plasmid-containing bacteria are selected by growth in the presence of tetracycline. The cloned DNA fragment is 1,000 bp long and has an EcoRI site 250 bp from one end. Three different recombinant plasmids are cleaved with EcoRI and analyzed by gel electrophoresis, giving the patterns shown below. What does each pattern say about the cloned DNA? Note: pBR322, the PstI and EcoRI restriction sites are about 750 bp apart. The entire plasmid with no cloned insert is 4,361 bp. Size markers in lane 4 have the number of nucleotides noted.

PCR errors during library amplification are one possible source of false positive results. If an error occurs in the first round of amplification, all the subsequent copies of that library fragment will also carry the variant, even though it is not present in the genome. However, reads from other library fragments spanning this same region will not have the variant, which means that identifying PCR copies, or clones, of library fragments during analysis can help to identify these types of errors. Based on what you know about PCR, which of the following statements would be true about the PCR clones in a sequencing library? A. PCR is subject to amplification bias, so reads derived from PCR clones will only map to regions that are not GC-rich. B. PCR is subject to amplification bias, so reads derived from PCR clones will only map to non-repetitive regions. C. PCR produces identical copies, so reads derived from PCR clones would map to the exact same location. D. PCR introduces many…

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Definition Definition Chemical process in which one or more monomers combine to produce a very large chain-like molecule called a polymer. The functional groups present on the monomers and their steric effects are responsible for polymerization through a sequence of reactions that vary in complexity. There exists a stable covalent chemical bond between monomers that sets apart polymerization from other processes.

Chapter 21, Problem 9EQ

Summary Introduction

To review:

The use of thermostable DNA (deoxyribonucleic acid) polymerase in PCR (polymerase chain reaction).

Introduction:

The technique that involves the cloning of the DNA fragment can either use vector (gene cloning) or work without it (PCR). In PCR, the DNA fragments are amplified using the dNTPs (deoxyribonucleotide triphosphate), polymerase, ligases, and primers.

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Answer 3

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Definition Definition Chemical process in which one or more monomers combine to produce a very large chain-like molecule called a polymer. The functional groups present on the monomers and their steric effects are responsible for polymerization through a sequence of reactions that vary in complexity. There exists a stable covalent chemical bond between monomers that sets apart polymerization from other processes.

Chapter 21, Problem 9EQ

Summary Introduction

To review:

The use of thermostable DNA (deoxyribonucleic acid) polymerase in PCR (polymerase chain reaction).

Introduction:

The technique that involves the cloning of the DNA fragment can either use vector (gene cloning) or work without it (PCR). In PCR, the DNA fragments are amplified using the dNTPs (deoxyribonucleotide triphosphate), polymerase, ligases, and primers.

Expert Solution & Answer

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Check out a sample textbook solution

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Answer 4

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Definition Definition Chemical process in which one or more monomers combine to produce a very large chain-like molecule called a polymer. The functional groups present on the monomers and their steric effects are responsible for polymerization through a sequence of reactions that vary in complexity. There exists a stable covalent chemical bond between monomers that sets apart polymerization from other processes.

Chapter 21, Problem 9EQ

Summary Introduction