Genetics: From Genes to Genomes
6th Edition
ISBN: 9781259700903
Author: Leland Hartwell Dr., Michael L. Goldberg Professor Dr., Janice Fischer, Leroy Hood Dr.
Publisher: McGraw-Hill Education
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Chapter 14, Problem 3P
Now that the sequence of the entire E. coli K12 strain genome (roughly 5 Mb) is known, you can determine exactly where a cloned fragment of DNA came from in the genome by sequencing a few bases and matching that data with genomic information.
| a. | About how many |
| b. | If you had purified a protein from E. coli cells, roughly how many amino acids of that protein would you need to know to establish which gene encoded the protein? |
| c. | You determine 100 nucleotides of sequence of genomic DNA from a different E. coli strain, but you cannot find a match in the E. coli K12 genome sequence. How is this possible? |
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Answer the following parts:
A. When performing classical Sanger or "dideoxy" sequencing, you set up 4 parallel reactions per template to be sequenced from a specific primer, with each of the four reactions containing a different dideoxynucleotide, and then the four reactions were run in a separate, adjacent lanes on a gel. Why couldn't you combine all 4 dideoxynucleotides with the primer and the template and do the whole reaction in one tube, and then run the set of fragments produced by the reaction mixture on a single lane in an acrylamide gel?
B. When doing automated sequencing, on the other hand, all 4 dideoxynucleotides are added to the same sequencing reaction, and run together in a single capillary gel. What's the difference - why can an automated sequencing reaction be done with all 4 dideoxynucleotides mixed together and all the resulting fragments run together, but not a conventional sequencing reaction?
Using the formulae for dsDNA to calculate g/mol:
You have a 4110 bp cloning vector
Want to ligate a 245 bp insert for molecular cloning.
efficient cloning requires that a 1:5 molar ratio of vector to insert is optimal for the ligation reaction.
What are the relative weights of vector and the insert DNA necessary in g?
A facility says they need 15 μL of a 40 ng/μL solution of plasmid DNA for sequencing. The typical yield for a DNA miniprep 5 μg eluted in 50 μL of solution. What do you need to do (for example dilution) to send the appropriate amount to the facility? Show all math work with an explanation.
Chapter 14 Solutions
Genetics: From Genes to Genomes
Ch. 14 - Choose the phrase from the right column that best...Ch. 14 - The unicellular, rod-shaped bacterium E. coli is 2...Ch. 14 - Now that the sequence of the entire E. coli K12...Ch. 14 - Bacterial genomes such as that of E. coli...Ch. 14 - List at least three features of eukaryotic genomes...Ch. 14 - Describe a mechanism by which a gene could move...Ch. 14 - High salt concentrations tend to cause protein...Ch. 14 - Recently, scientists tested the possibility that...Ch. 14 - A recent metagenomic study analyzed the...Ch. 14 - Linezolid is a new type of antibiotic that...
Ch. 14 - A liquid culture of E. coli at a concentration of...Ch. 14 - Pick out the medium i, ii, iii, or iv onto which...Ch. 14 - This problem concerns Fig. 14.14, which...Ch. 14 - In two isolates one is resistant to ampicillin,...Ch. 14 - E. coli cells usually have only one copy of the F...Ch. 14 - In E. coli, the genes purC and pyrB are located...Ch. 14 - DNA sequencing of the entire H. influenzae genome...Ch. 14 - Genes encoding toxins are often located on...Ch. 14 - a. You want to perform an interrupted-mating...Ch. 14 - In Problem 19, do you think that most of the...Ch. 14 - One issue with interrupted-mating experiments such...Ch. 14 - Prob. 22PCh. 14 - Starting with an F- strain that was prototrophic...Ch. 14 - You can carry out matings between an Hfr and F...Ch. 14 - Genome sequences show that some pathogenic...Ch. 14 - Generalized and specialized transduction both...Ch. 14 - This problem highlights some useful variations of...Ch. 14 - A researcher has a Trp auxotrophic strain of E....Ch. 14 - Streptococcus parasanguis is a bacterial species...Ch. 14 - The sequence at one end of one strand of the...Ch. 14 - Scientists who study amino acid biosynthesis...Ch. 14 - Suppose that you could obtain radioactively...Ch. 14 - Prob. 34PCh. 14 - Some scientists are trying to engineer...
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- For each scenario below, pick which technique you might use to answer the questiion 1. You have received a new shipment of DNA from a collaborator and need to determine the concentration of the DNA before using it. 2. You have identified a mutant in the oxygen carrying protein, Hemoglobin, which you believe alters the protein from forming a tetramer. You want to determine whether it forms a tetramer and if the overall structure has changed. (Probably need two different techniques here) 3. You want to determine whether the protein I-contain-tryptophan2 is unfolding in solution with the addition of a drug-like molecule. 4. You have identified the sequence of a new gene, but are unsure of what the resulting product might look like and if it would be a viable protein-coding genearrow_forwardE. How many nucleotides would be required to generate a polypeptide that is 15 amino acids long? This requires knowing how many nucleotides of DNA code for one amino acid. F. Assuming that there are between 20,000-25,000 genes in the human genome, do you think there are 1) fewer, 2) approximately the same number, or 3) more proteins in the human genome? Explain your answer.arrow_forwardWe are given a random DNA sequence (each of the 4 bases has equal probability of occurring at each position in the sequence). A. What is the probability that three adjacent bases (a triplet) encode a stop codon? B. What is the probability that a given triplet is not a stop codon? C. What is the probability that neither of two adjacent triplets is a stop codon? D. What is the probability that none of 100 adjacent triplets are stop codons? E. Consider a single base insertion mutation between the 3rd and 4th codons in a natural gene that encodes a protein 100 amino acids long. Is it more likely that the protein produced by this mutant allele will be shorter or longer than 100 amino acids long?arrow_forward
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