1. What is the method for isolating plasmid DNA in E. coli? 2. What is the metho
ID: 280606 • Letter: 1
Question
1. What is the method for isolating plasmid DNA in E. coli? 2. What is the method for isolating total genomic DNA from Drosophila? 3. Why precipitate genomic DNA but do no precipitate the other? 4. If isolating total genomic DNA from mammalian tissue culture cells, could you use the same method as that of Drosophila? What changes might you make? 5. What is the purpose of solution 2 (SDS and NaOH) in the plasmid DNA extraction procedure? Why isn't it included in Drosophila genomic DNA isolation method? 6. If a Jr. Scientist used the plasmid DNA purification method with adult Drosophila, what would be the most likely outcome of the PCR amplification experiment?Explanation / Answer
Answer 1: For the isolation of plasmid DNA from E.coli, the alkaline lysis method is most suitable method which is invented by Birnboim and Doly (1979).
Reference:
Birnboim, H. C. and Doly, J. (1979). A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7(6): 1513-1523.
https://bio-protocol.org/bio101/e30#biaoti20063
Answer 2. The Phenol - chloroform DNA extraction method can be used for isolating the total genomic DNA from Drosophilla as described by Sambrook and Russel (2001).
Reference:
Commonly Used Techniques in Molecular Cloning, Appendix 8, in Molecular Cloning, Volume 3, 3rd edition, by Joseph Sambrook and David W. Russell. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2001.
http://cshprotocols.cshlp.org/content/2006/1/pdb.prot4455.long
Answer 3. The DNA or Nucleic acid is hydrophillic in nature which is dissolved in water and when the sodium acetate is added followed by ethanol and decrease the temperature converts DNA molecule in to less hydrophillic and leads to precipitates and drops out from solution, where as other molecules remain disolved in it.
Reference:
https://bitesizebio.com/253/the-basics-how-ethanol-precipitation-of-dna-and-rna-works/
Answer 4: For the isolation of geneomic DNA from Drosophila and mammalian tissue, we can use the same protocol ie. the Phenol - chloroform DNA extraction method with minor modification as described by Sambrook and Russel (2001).
Reference:
Commonly Used Techniques in Molecular Cloning, Appendix 8, in Molecular Cloning, Volume 3, 3rd edition, by Joseph Sambrook and David W. Russell. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2001.
https://www.qiagen.com/us/resources/molecular-biology-methods/dna/
Related Questions
Navigate
Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.