I have the cDNA for the KIF3A gene in circular, double-stranded DNA plasmid in E
ID: 34756 • Letter: I
Question
I have the cDNA for the KIF3A gene in circular, double-stranded DNA plasmid in
E. Coli. I want to sequence the first five bases to confirm that I have the proper gene and that
there are no mutations.
a) Normally you would use a 15-mer oligonucleotide primer to sequence the gene, what
are the last five bases of the primer you will use?
b) What are the 5 crucial ingredients in your sequencing reaction?
c) When the results of the sequencing reaction are run out on a gel, there will be bands
corresponding to different length sequences. From top to bottom of the gel, give the sequence of
the DNA that has been added to the sequencing primer for the 5 bases we are interested in
sequencing.
Explanation / Answer
KIF3A - kinesin like protein thait is encoded by KIF3A GENE ,it is heterodimeric protein consists of kap3 protien complex. it ios present in cilium ,cytosol and cytoskeleton.
cDNA is the complementary dna which is produced using reverse transcriptase enzyme.
a. 5?-CAGGCCAACAGGAGCAAACAT-3?