Question

3. Put the following steps in order (#1-7) to illustrate the experimental design flow for our molecular module analysis of yellow gene sequences from wildtype and mutant flies: (3pt)

   • Isolate plasmid with region of interest from bacteria

   • Isolate gDNA

   • Sequence region of interest from plasmid

   • Insert region of interest into plasmid

   • Amplify gDNA region of interest

   • Put plasmid with region of interest into bacteria to copy

   • Digest plasmid to verify region of interest is present

Answer 1

1) Isolate gDNA .

2) amplify gDNA region of interest .

3) Isolate plasmid with region of interest from bacteria.

4) sequence region of interest from plasmid.

5) digest plasmid to verified region of interest is present.

6) insert region of interest into plasmid.

7) put plasmid with reagion of interest into bacteria to copy.

Summary :- plasmid is the extrachromosmal DNA that is present in the some bacteria . It is round in shape . It has the property of replication independent of the the replication in the nucleus. The plasmid is capable of producing large number of copies. The gene of interest is isolated by the process of gel electrophoresis. Firstly the gene of interest is cut by endonuclease and exonuclease. The plasmid is also cut through the same and nucleus and its nucleus this result in the same origin of replication. The gene of interest is then inserted into the plasmid with the help of DNA ligase. This plasmid molecule is an inserted into host cell in which it replicate at a higher rate. This process is mainly used to obtain certain proteins such as insulin .

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