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Publisher Summary This chapter discusses one-sided anchored polymerase chain reaction (PCR) for the amplification and sequencing of complementary DNA (cDNA). It presents a modified PCR protocol, anchored PCR that allows the amplification of a specific full-length messenger RNA (mRNA) when only a small amount of sequence information is available. The two rounds of anchored PCR amplification result in a single homogeneous amplified product that can then be directly sequenced or cloned into an appropriate vector for further analysis. A modification of this method has also been used to detect transcription initiation sites in the phospholipase A2 gene. The sensitivity of this method for detecting transcription initiation may be 10-fold greater than that of conventional primer extension approaches, making it useful for the analysis of rare mRNA species. As with any method for the analysis of mRNA, the success of these protocols depends on the quality and integrity of the mRNA being used and on the successful synthesis of cDNA templates. Appropriate precautions to ensure a high yield of intact mRNA and full-length cDNA should be taken.