a. If I wanted to generate a PCR product amplifying the region between exon 1 an

Question

a. If I wanted to generate a PCR product amplifying the region between exon 1 and exon 3, what would be the sequence of the two oligonucleotide primers that I would have to use (for simplicity, each primer can be just 6nt long) ? Be sure to indicate the 5’ and 3’ ends of each primer.

b. Having created the PCR primers, what would be the size of the product if I used genomic DNA as the template for PCR? Please include the size of your primers in the product.

c. What would be the size of the product if I used cDNA as the template for PCR?

d. Some individuals in the population have a 300-bp Alu element inserted into intron 1, at the location indicated by an X, that causes a recessive form of hemophilia. How would your responses in b and c change if the template nucleic acid were from an individual homozygous for the allele with the inserted Alu element?

1. The gene structure of the human X-linked clotting factor, Factor XIV, is shown below. The gene comprises three exons and two introns. The sizes of exon 2, and of the two introns, is indicated. Also, a small amount of sequence from the coding strand portion of exons 1 and 3 is shown. These sequences read 5' to 3', left to right. CTGGAC AGATTC 2 3 800bp 200bp 400bp

Explanation / Answer

Answer

Reverse primer 5’-GTCCAG-3’

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