Genetic Analysis: An Integrated Approach (3rd Edition)
3rd Edition
ISBN: 9780134605173
Author: Mark F. Sanders, John L. Bowman
Publisher: PEARSON
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Chapter 16, Problem 8P
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Transcriptome analysis involves two separate methodologies: gene expression and RNA seq analyses. The 10 items below are a scrambled listing of the steps used in the two procedures. Identify the steps involved in RNA seq from the list below. Use the numbers in the list to refer to each step. Once the steps for RNA seq have been identified, write the steps in the order in which they are performed during the experiment.
(1) DNA sequencing
(2) Allow for hybridization and wash excess cRNA.
(3) Mix labeled cRNA with array chip.
(4) PCR amplification
(5) Measure fluorescence intensity to determine abundance of transcripts.
(6) Add labeled cRNA at each microarray location.
(7) Map cDNA sequences to the genome of the organism to determine identity and abundance of transcripts.
(8) mRNA isolation from cells
(9) Prepare fluorescently labeled cRNA probes
(10) cDNA synthesis
You just graduated from college and started working at a biotech startup called Scrofabulous. Your
first job assignment is to clone the pig gene for the hormone prolactin. Assume that the pig gene
for prolactin has not yet been isolated, sequenced, or mapped; What would be the most useful and
economical first step to go about identifying and cloning the pig gene for prolactin?
use the amino acid sequence of mouse prolactin to design a pair of degenerate oligonucleotide PCR primers
to PCR-amplify the pig prolactin gene.
RNAseq the pituitary gland of the pig, the most abundant gene is likely to to be prolactin
Conduct a proteome search for peptides that match parts of mouse prolactin protein
Sequence the pig genome, then translate the genome to find the gene predicted to encode for prolactin
Crystalize the mouse prolactin protein and use Google's DeepMind Al to find the closest amino acid
sequence in the pig proteome
what is the whole-genome shotgun sequencing? Also briefly explain its strategy to assemble the genome sequence.
Chapter 16 Solutions
Genetic Analysis: An Integrated Approach (3rd Edition)
Ch. 16 - You have discovered a new species of Archaea from...Ch. 16 - 16.2 Repetitive DNA poses problems for genome...Ch. 16 - 16.3 When the whole-genome shotgun sequence of the...Ch. 16 - How do cDNA sequences facilitate gene annotation?...Ch. 16 - 16.5 How do comparisons between genomes of related...Ch. 16 - 16.6 You are designing algorithms for the...Ch. 16 - 16.7 You have sequenced a region of the Bacillus...Ch. 16 - You have just obtained 100-kb of genomic sequence...Ch. 16 - 16.9 The human genome contains a large number of...Ch. 16 - Based on the tree of life in Figure 16.12, would...
Ch. 16 - 16.11 When comparing genes from two sequenced...Ch. 16 - 16.12 What is a reference genome? How can it be...Ch. 16 - Prob. 13PCh. 16 - Prob. 14PCh. 16 - 16.16 Consider the phylogenetic tree below with...Ch. 16 - You have isolated a gene that is important for the...Ch. 16 - 16.18 When the human genome is examined, the...Ch. 16 - Symbiodinium minutum is a dinoflagellate with a...Ch. 16 - Substantial fractions of the genomes of many...Ch. 16 - 16.21 A modification of the system, called the ...Ch. 16 - 16.22 A substantial fraction of almost every...Ch. 16 - 16.23 In the globin gene family shown in Figure ,...Ch. 16 - You are studying similarities and differences in...Ch. 16 - In conducting the study described in Problem 24,...Ch. 16 - Prob. 26PCh. 16 - Prob. 27PCh. 16 - Prob. 28PCh. 16 - Prob. 29P
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- Consider the following distance matrix: A B C D A B C D ☆ 0.1 0.4 0.3 0.8 0.6 0.2 Using the UPGMA algorithm, answer the following questions: a) What two sequences are the most related? Which are the least related? b) Does the following tree accurately capture the relationship among the sequ in the distance matrix (ignoring branch lengths)? Explain in detail why or whyarrow_forwardIf you have access to the necessary computer software, make asequence file and analyze it in the following ways: What is thetranslated sequence in all three reading frames? What is the longest open reading frame? Is the sequence homologous to any known sequences? If so, does this provide any clues about the function of the sequence?arrow_forwardWhy are closure and completeness important in genome sequencing?arrow_forward
- a. What type of nucleic acid and from what species would the scientist use to begin construction of her genomic DNA library? b. From what tissue would she isolate this nucleic acid? c. What type of reagent would the scientist use to cut the genome into appropriately sized fragments? d. What size nucleic acid fragments would one aim to prepare for the library construction so as to to avoid having to screen an overwhelming number of clones? e. Into what vector would the scientist ligate her genomic DNA fragments? f. What organism would the scientist use to propagate the clones of her genomic DNA library? g. From the information given in the problem determine what probe could be used to screen the scientist's library to find her clone of interest ?arrow_forwardDescribe the difference between Sanger based sequencing and Next Generation Sequencing (NGS). Why is NGS advantageous over Sanger based sequencing?arrow_forwardWhat does the symbol “N” indicate (see the arrow)? Is this a problem for getting an accurate DNA sequence? The height of the peak in the figure is an indication of the quality of the data, with a high peak correlated with a low error in “base-calling” of perhaps 1 error in 10,000 bases. Is this generally a high-quality DNA sequence for determining the true DNA sequence of the gene studied? How can you tell? Which end of the sequence shown above is the 5’ end of the sequence being made via PCR.arrow_forward
- What provides a convenient bridge between the low resolution of a karyotype and the ultra-high resolution of a complete genomic sequence?arrow_forwardWhole-exome sequencing (WES) is helping physicians diagnose a genetic condition that has defied diagnosis by traditional means. The implication here is that exons in the nuclear genome are sequenced in the hopes that, by comparison with the genomes of nonaffected individuals, a diagnosis might be revealed. (a) What are the strengths and weaknesses of this approach? (b) If you were ordering WES for a patient, would you also include an analysis of the patient’s mitochondrial genome?arrow_forwardWhile comparative genomics is fundamentally the study of the differences between the genomes of species, it can also give clues about functional genomics (the study of the biological functions that rely on genomic sequences i.e. related sequences that encode the various proteins within a family). How? Give an examplearrow_forward
- What is a repetitive element in genomics? What are the types of repetitive elements? What is their effect on the ease of determining and analyzing a genome sequence?arrow_forwardIn the practical you have been analysing a human genomic library. You know from your calculations that only a small proportion of the human genome is represented, even when the entire class results are considered. Therefore, the chance of finding a particular single-copy gene in your library is very small. Outline a strategy for constructing a genomic DNA library more representative of the entire human genome. You will need to consider alternative vectors and the efficiency of transformation of the bacterial cells.arrow_forwardWhat is the primary disadvantage of Sanger sequencing?arrow_forward
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Genome Annotation, Sequence Conventions and Reading Frames; Author: Loren Launen;https://www.youtube.com/watch?v=MWvYgGyqVys;License: Standard Youtube License