1. A researcher plans to amplify the CDS of the SAM1 gene. The partial sequence below shows the coding sequences for the N-terminus and C-terminus of Sam1p. Design two 18-nucleotide long primers to amplify the sequence the SAM1 CDS.
5’-ATGGCCGGTACATTTTTATTC..........(CDS).......TCCAAGACTTTGAAGTTCTAA - 3’
2. Most of the primers that we are using for PCR have Tms that are slightly lower than the annealing temperature used in the PCR reactions. Thus, less than half of the target sites are expected to anneal with the primers in each cycle. Why would investigators choose to design primers that will not be fully annealed with the DNA template?