24. You discover a novel EUKARYOTIC organism that glows in the dark. You believe

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24. You discover a novel EUKARYOTIC organism that glows in the dark. You believe this trait is due o a single gene, and you wish to clone the gene. Which of the following strategies is most likely to be successful? a. Iso late the genomic DNA from the organism, digest with a restriction endonuclease, insert into a plasmid vector and transform into bacteriA. Screen colonies for the ability to glow in the dark Isolate the genomic DNA from the organism, digest with a restriction endonuclease, insert into a plasmid vector and transform into eukaryotic cells such as yeast. Screen colonies for the ability to glow in the dark Isolate mRNA from the organism, reverse transcribe and generate cDNA, insert into a plasmid vector and transform into bacteriA. Screen colonies for the ability to glow in the dark Isolate mRNA from the organism, reverse transcribe and generate cDNA, insert into a plasmid vector and transform into eukaryotic cells such as yeast.Screen colonies for the ability to glow in the dark b. c. d. 25. Which of the following would you expect to be a drawback to PCR techniques? PCR has the potential to create positive results from very low levels of contamination a. b. PCR can be used with very small samples such as single sperm cells C. PCR is very rapid; in some cases results can be available in a few hours. d. All of these. 26. In choosing a vector, viral vectors typically can carry much larger pieces of DNA than plasmid vectors but plasmid vectors are much easier to work with since they do not kill their host cells. In which of the following situations would you anticipate preferring a viral vector? a. You need to clone an entire gene, including regulatory sequences b. You are generating a genomic library from an organism with a large genome. C. You are creating an mRNA library to identify the gene encoding a very large protein. d. All of these e. None of these 27. Electroporation is: a. b. c. d. the process of separating charged molecules through a gel maintained in an electric field the process of combining foreign DNA to an electrically charged vector molecule the process of introducing DNA into cells by the application of high voltage pulses All of the above

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Ans 24)

Ans d) Isolate mRNA from the organism, reverse transcribe and generate cDNA, insert into a plasmid vector and transform into eukaryotic cells such as yeast. Screen colonies from the ability to glow in the dark.

In this case, while using this process, it is possible to actually screen large number of genomic DNA which is present in the eukaryotes.

Ans 25) a) PCR has the potential to create positive results from very low levels of contamination.

This is a major drawback because it might lead to wrong results and inappropriate date interpretation.

Ans 26) d) All of these

In all of the situations we prefer only a viral vector and it helps in better cloning process and creation of libraries.

Ans 27) d) All of the above

Electroporation is a microbial technique which uses an electrical field which is applied to the cells and it increases the overall cell membrane permeability. This enables the chemicals, drugs and DNA to be introduced into the cell

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