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Scheme of 5′ SAGE methodology. The poly(A)-rich RNA is divided into two pools. Different oligos set (blue or red) are used to carry out reverse transcription, template-switching and primer extension. After tagging enzyme (yellow oval) digestion, the two sample pools are combined together to make 120 bp ditags. The anchoring enzyme (blue triangle) is used to generate 50 bp ditag for concatenation. Specifics are discussed in Materials and Methods.
