Cdna sequence and pcr

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Reference no: EM1384462

You want to fuse a cDNA sequence to the end of two tandem"HA" antibody epitope tags within the MCS-HAHA (multiple cloning site-HAHA) region of the vector shown below. When transformed into yeast this plasmid will express the fusion protein under the transcriptional control of the pGAL1 promoter as shown. The corresponding sequence of the MCS-HAHA region is shown from the EcoRI site to the Sal1 site, and the reading frame of the two HA tags is indicated (each HA epitope is designated by the underlined amino acid sequence). Design primers with XhoI and SalI sites for PCR amplification of the entire cDNA (sequence at the bottom) to clone it together, in frame, with the HAHA epitope in the vector. Calculate for both primers their respective Tm and report the sequence of temperatures and times required for one cycle of the PCR amplification.

Reference no: EM1384462

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