Question

RT-PCR involves a series of two distinct enzymatic reactions. For the RT step what is the enzyme and what is the substrate? For the PCR step, what is the enzyme and what is the substrate? 3. Is the product of these experiments single or double stranded? 4. How will we use gel electrophoresis in these experiments?

Answer 1

For the RT step what is the enzyme and what is the substrate?
The first step in RT PCR is reverse transcription => Sunthesis of DNA from mRNA. Reverse transcriptase catalyzes this first step and the substrate is mRNA.

For the PCR step, what is the enzyme and what is the substrate?
For the PCR step, the DNA polymease catalyzes this step and the substrate is cDNA.

3. Is the product of these experiments single or double stranded?
The prodcuts of this experiment is double stranded DNA.

4. How will we use gel electrophoresis in these experiments?
To get a clear resluts, to avoid the RNA contamination sometimes we use RNase treatment to amplified PCR prodcuts the, load that PCR sample in the gel along with loading dye. So we can mesure the amplifed fragment length.