Understanding qRT-PCR exercise
Name:____________________
In the gene sequence provided, the exons are labeled as uppercase and the introns labeled as
lowercase including the consensus splice sites at the beginning (gt) and end (ag) of the intron.
1. Using the provided gene sequences, create the corresponding cDNA sequences (which
represents the mRNA sequences) . Paste here
2. The position of the two primers used during the RT-PCR set up from last week is
indicated by underline. Calculate the expected size of the amplified RT-PCR fragment.
Remember that RT-PCR is a 2-steps protocol; 1st step is converting the RNA into cDNA,
second step is amplifying sequence between the two primers.
Expected size for your RT-PCR reaction (in bp): _________________________
3. Using the ExPasy Translate tools obtain the corresponding protein sequence from the
cDNA. Paste here
4. Use the protein sequence to identify protein function and homologs in other species
using NCBI/BLASTp. Provide a screen shot showing
a. the first two parts of the BLAST result page
b. one of the alignments.