Abstract
Genomic amplification with transcript sequencing (GAWTS) (Stoflet et al., 1988; Sommer et al., 1990) is a generally applicable method for direct sequencing of PCR material. GAWTS is centered around the attachment of a phage promoter sequence (T7, SP6, or T3) to the 5′-end of one or both PCR primers. The phage promoter sequence allows the PCR product to be transcribed into RNA. Subsequently, the RNA is utilized as a single-stranded template for dideoxynucleotide sequencing with reverse transcriptase (Figure 18.1).
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Sommer, S.S., Vielhaber, E.L. (1994). Phage Promoter-Based Methods for Sequencing and Screening for Mutations. In: Mullis, K.B., Ferré, F., Gibbs, R.A. (eds) The Polymerase Chain Reaction. Birkhäuser, Boston, MA. https://doi.org/10.1007/978-1-4612-0257-8_18
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DOI: https://doi.org/10.1007/978-1-4612-0257-8_18
Publisher Name: Birkhäuser, Boston, MA
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