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==Primer Design== *I received this pair of primers from Sergio CGG(RE)ATGGAAGAGGATCAGGAGCTGGAGAG (Forward) 5β-3β CGG(RE)CTAGAATGTGGATTCATTTCTGCTTGC (Reverse) 5β-3β *I am not going to add restriction enzyme, and hence I will just order the primers without the RE sites. *After blasting the primers o NCBI blast the the primers matched 100% to POLR1D gene when cgg is removed from 5' end *Order from IDT after confirming with Kun ATGGAAGAGGATCAGGAGCTGGAGAG (Forward) 5β-3β CTAGAATGTGGATTCATTTCTGCTTGC (Reverse) 5β-3β *After blasting the primers aligns at transcript NM_015972.3 this locus is 817bp long, with primer 3 the product size is 402 with Tm at 67.46C and 63.40C (avg~65C)for forward and reverse primer respectively. I will use annealing temperature at 55C
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