Given a large section of a genome say a complete chromosome

Given a large section of a genome, say a complete chromosome, how could you locate genes? Given a large section of a gene, say a complete exon, how could you determine reading frame?

Solution

3. The following steps you should follow to isolate a required gene sequence in chromosome:

1.      Designing primers against the known sequence of the target DNA you want to locate (forward and reverse primer)

2.      Putting PCR (polymerase chain reaction) reaction with necessary chemicals and temperature profile by using DNA as template (given chromosome).

3.      Running the PCR product in agarose by agarose gel electrophoresis by using a specific ladder according to the your product size

4.      Locating the DNA product in UV-transilluminator and cutting the gel where your DNA of interest is located

5.      Elution of the DNA from the gel

6.      Eluted product will contain your DNA of interest segment

4. The following steps you should follow to isolate a required exon (gene) sequence:

Exon portion of the gene is located in mRNA sequence only

1.      RNA isolation

2.      cDNA synthesis

3.      Primer designing on the bases of RNA sequence of the required gene

4.      Putting PCR (polymerase chain reaction) reaction with necessary chemicals and temperature profile by taking cDNA as a template.

5.      Running the PCR product in agarose by agarose gel electrophoresis by using a specific ladder according to the your product size

6.      Locating the DNA product in UV-transilluminator and cutting the gel where your DNA of interest is located

7.      Elution of the DNA from the gel

8.      Eluted product will contain your DNA of interest segment