Background: You are working in a biochemistry/molecular biology lab that special
ID: 146076 • Letter: B
Question
Background: You are working in a biochemistry/molecular biology lab that specializes in protein applications (purification, identification, etc). The lab has been working with a new organism. This organism is single-cellular and eukaryotic and appears to be related to yeast. You are entering the lab and have been assigned characterizing this new organism.
Isolating and Characterizing Nucleic Acids 6. How might you use genomic DNA from this organism to compare to other yeast organisms? a. how would you manipulate the DNA (restriction enzymes)? b. What is a southern blot useful for? c. How would you sequence it and compare? (saving for second test block)Explanation / Answer
Ans a) Genetic manipulation is a technique through which one can actually manipulate the DNA with help of different factors and things. The process of DNA manipulation basically requires the cleavage which is done by restriction enzymes. The isolation of DNA can be done where the DNA strands are broken with the restriction endonucleases on the specific site. This is followed by insertion of the gene of interest and joining the DNA through the process of ligation. The genomic DNA manipulation majorly uses different type of restriction enzymes for the process.
Ans b) The Southern blot is a method which helps in detecting a specific sequence of the DNA is an overall DNA sample. It combines the electrophoresis method to separate the DNA fragments and identity it as separate entity. It basically uses the methods of gel electrophoresis and hydridization to analyze the specific sequences of DNA. During the electrophoresis, the DNA fragments are separated based on their size and charge and helps it becomes easy to detect the DNA using a specific DNA probe.
Ans c) The sequences of the DNA are compared to each other through a number of processes. It is basically established through the sequence homology with the concept that sequences have evolved from a common ancestor. It is the aligning of the sequences and finding out for the homology through techniques like BLAST and FASTA
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