Why are TOPO vectors used for cloning PCR products? What is the efficiency of TO

Question

Why are TOPO vectors used for cloning PCR products? What is the efficiency of TOPO cloning over ligation?

How are Lambda () viruses modified to make useful cloning vectors? Describe how cloning vectors and insert DNA are prepared to carry out cloning. What advantages do -based vectors have over plasmid vectors?

Compare plasmid and -based vectors to cosmid vectors, what are the advantages of cosmid vectors? What features would you expect to find in a Yeast Artificial Chromosome, what is their purpose?

What is the advantage of BACs over YACs?

Give the approximate insert size accepted by the various cloning vectors?

What are the various types of cloning libraries? What is the advantage/use of each type? How is the source material prepared for each type of library?

What variables determine the number of clones needed to get reasonable coverage in a genomic library?

In screening various libraries by colony or plaque hybridization what are some likely sources for your probes?

Describe how differential screening can be used to find cell type specific clones. How is chromosome walking used to locate a gene sequence?

What is an ordered library? How can it be efficiently screened?

Describe how transposon tagging can be used to find/clone a gene. What is rescue cloning in relation to transposon tagging?

Explanation / Answer

1.

TOPO cloning, Topoisomerase based cloning is a method used for cloning DNA and this method do not use any ligases or restriction enzymes. TOPO reaction works using the intrinsic properties of the enzyme topoisomerase. This enzyme ligates the vector and the insert within a matter of minutes. This method is very much efficient and results in 95% successful rate.

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