Essentials Of Genetics (9th Global Edition)
9th Edition
ISBN: 9780134143637
Author: William S. Klug, Michael R. Cummings
Publisher: Pearson Global Edition
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Chapter 17, Problem 15PDQ
Summary Introduction
To review:
In cDNA library formation, shorter cDNA clones are derived as compared to mRNA molecules.
Introduction:
cDNA Libraries are a collection of cDNA of one organism cloned into host cells. They are formed from mRNA isolated from cells. They only contain coding sequences and do not contain introns or other non-coding regions. Due to this reasons, cDNA libraries are preferred over genomic libraries. cDNA libraries are created by inserting vectors into cDNA and cloning to produce more copies.
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What advantages do cDNA libraries provide over genomic DNA libraries? Describe cloning applications where the use of a genomic library is necessary to provide information that a cDNA library cannot.
After Create a second cDNA strand complementary to the first. After the reaction is completed, the enzyme S1 nuclease is used to cleave the hairpin loop., what is the final step of making a cDNA library?
One procedure of obtaining cDNA from mRNA is by using oligo(dT) primers. What are oligo(dT)s? Why does using them make sense based on the processing (or modification) of precursor mRNA to get mature mRNA?
Chapter 17 Solutions
Essentials Of Genetics (9th Global Edition)
Ch. 17 -
CASE STUDY |Should we worry about recombinant DNA...Ch. 17 - Prob. 2CSCh. 17 - Prob. 3CSCh. 17 -
HOW DO WE KNOW?
1. In this chapter we focused on...Ch. 17 - Prob. 2PDQCh. 17 - What roles do restriction enzymes, vectors, and...Ch. 17 - Prob. 4PDQCh. 17 - Prob. 5PDQCh. 17 - Prob. 6PDQCh. 17 - Prob. 7PDQ
Ch. 17 - List the advantages and disadvantages of using...Ch. 17 - What are the advantages of using a restriction...Ch. 17 - The introduction of genes into plants is a common...Ch. 17 - Prob. 11PDQCh. 17 - Prob. 12PDQCh. 17 - Prob. 13PDQCh. 17 - What advantages do cDNA libraries provide over...Ch. 17 - Prob. 15PDQCh. 17 -
16. List the steps involved in screening a...Ch. 17 -
17. In a typical PCR reaction, describe what is...Ch. 17 -
18. We usually think of enzymes as being most...Ch. 17 - How are dideoxynucleotides (ddNTPs) structurally...Ch. 17 - Prob. 20PDQCh. 17 - One complication of making a transgenic animal is...Ch. 17 -
22. When disrupting a mouse gene by knockout, why...Ch. 17 - Prob. 23PDQ
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- What is the advantage of using a cDNA library?arrow_forwardIf you wanted to express a library of human proteins in yeast, there are several good reasons why it would be better to use a cDNA library instead of a genomic library from humans. Which one of the following options is not such a reason? a. yeast may not initiate transcription or splicing of a human gene at the correct locations b. some genes will have very high representation (many plasmids contain the gene), while others will have very low representation (few or no plasmids contain the gene) in the cDNA library c. most genomic library clones would be useless, because only ~1.5% of the human genome actually encodes proteins d. a cDNA library can contain multiple splice variants, which are common for human genes e. introns are often very large in the human genome, making it impossible in many cases to contain a genomic version of an entire gene in a single plasmidarrow_forwardWhen a region of DNA that contains the genetic information for a protein is isolated from a bacterial cell and inserted into a eukaryotic cell in a proper position between a promoter and a terminator, the resulting cell usually produces the correct protein. But when the experiment is done in the reverse direction (eukaryotic DNA into a bacterial cell), the correct protein is often not produced. Can you suggest an explanation?arrow_forward
- a. What sequence information about a gene is lackingin a cDNA library?b. Can clones in a cDNA library contain 5′ UTR sequences? 3′ UTR sequences?c. Would you be likely to find on average longerORFs in cloned sequences from a genomic libraryor from a cDNA library? Explainarrow_forwardA protein has the following amino acid sequence: Met-Tyr-Asn-Val-Arg-Val-Tyr-Lys-Ala-Lys-Trp-Leu-Ile-His-Thr-Pro You wish to make a set of probes to screen a cDNA library for the sequence that encodes this protein. Your probes should be at least 18 nucleotides in length. Q. How many different probes must be synthesized to be certain that you will find the cDNA sequence that specifies the protein?arrow_forwardYou isolate a mouse Tau-gene-containing DNA fragment from the chicken and hybridize it to the freshly-made and isolated hnRNA (primary transcript) from the nucleus of the mouse cells transcribed from the Tau gene (immediately after it was produced), allowing no time for processing of the hnRNA. Describe what you see when you look at the DNA/RNA hybrid molecule under the electron microscope.arrow_forward
- After Drosophila DNA has been treated with a restriction enzyme, the fragments are inserted into plasmids and selected as clones in E. coli. With the use of this “shotgun” technique, every DNA sequence of Drosophila in a library can be recovered.a. How would you identify a clone that contains DNA encoding the protein actin, whose amino acid sequence is known?b. How would you identify a clone encoding a specific tRNA?arrow_forwardCould quantitative PCR, which uses a DNA-binding dye, to show how many copies of the target DNA sequence could be used to quantify the amount of mRNA in a cell? Would you expect that a metabolically active tissue such as the liver would show more cDNA copies in such a method, compared to less metabolically active tissues such as skin cells? One reason that the types and amounts of mRNAs are quantified in different tissue types is to compare which genes are activated and which are inactive. It used to be thought that any gene that was transcribed was automatically translated. The discovery of RNA-degrading systems shows that the real situation in cells is more complemented. Do you believe that a larger amount of mRNA of a given type, say for alpha hemoglobin in immature red blood cells is a reliable indicator that more alpha hemoglobin protein will be made in those cells?arrow_forwardWhat is the difference between a genomic library and a cDNA library?arrow_forward
- Suppose you had isolated a new transcription factor and wanted to know which genes this protein might regulate. Is there any way that you could use a cDNA microarrayarrow_forwardCreate a hybrid cDNA-mRNA molecule using reverse transcriptase and oligo-dT primer, what is step 3 of making a cDNA library?arrow_forwardDescribe the difference between a transcriptional fusion and a translational reporter gene fusion. What sequences should you include in each? What types of information can be garnered from their analyses? Please include annotated pictures of the reporter gene expression patterns.arrow_forward
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