Fig 8. Part of the cDNAs cloned via a primer annealing only at 4 or 5 nts during PCR.
Top panel: a cDNA was inadvertently cloned because the last 4 nts, i.e. 5’GGAC3’ (shaded), of our mtR3071 primer (underlined and boldfaced) are reversely complementary and thus annealed to the 2263-2266th bps of the mtDNA (AY195786.2). The lowercase letter “t” at the 5’ side of the mtR3071 sequence was appended by Taq DNA polymerase, and the sequence before this “t” belongs to the T-A vector. Bottom panel: A cDNA was accidently cloned because the last 5 nts, i.e. 5’TCTTC3’ (shaded) in the mtR2694 primer (underlined and boldfaced), is reversely complementary and thus mistakenly annealed to the 3550-3554th-bp region of the mtDNA (AY195786.2). The lowercase letter “a” after the primer was appended by Taq DNA polymerase, whereas the sequence after the “a” belongs to the T-A vector. (“…” sequence omitted)