Chapter 20: Problem 18
List the steps involved in screening a genomic library. What must be known before starting such a procedure? What are the potential problems with such a procedure, and how can they be overcome or minimized?
Chapter 20: Problem 18
List the steps involved in screening a genomic library. What must be known before starting such a procedure? What are the potential problems with such a procedure, and how can they be overcome or minimized?
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Get started for freeTo create a cDNA library, cDNA can be inserted into vectors and cloned. In the analysis of \(\mathrm{cDNA}\) clones, it is often difficult to find clones that are full length-that is, many clones are shorter than the mature mRNA molecules from which they are derived. Why is this so?
What is the difference between a knockout animal and a transgenic animal?
In a control experiment, a plasmid containing a HindIII recognition sequence within a kanamycin resistance gene is cut with HindIII, re-ligated, and used to transform \(E\). coli \(\mathrm{K} 12\) cells. Kanamycin-resistant colonies are sclected, and plasmid DNA from these colonies is subjected to electrophoresis. Most of the colonies contain plasmids that produce single bands that migrate at the same rate as the original intact plasmid. A few colonies, however, produce two bands, one of original size and one that migrates much higher in the gel. Diagram the origin of this slow band as a product of ligation.
How do next-generation sequencing (NGS) and third. generation sequencing (TGS) differ from Sanger sequencing?
Assume you have conducted a DNA sequencing reaction using the chain- termination (Sanger) method. You performed all the steps correctly and electrophoresced the resulting DNA fragments correctly, but when you looked at the sequencing gel, many of the bands were duplicated (in terms of length) in other lanes. What might have happened?
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