Genomic Amplification with Transcript Sequencing (GAWTS)

  • Tammy Lind
  • Erik C. Thorland
  • Steve S. Sommer
Part of the Methods in Molecular Biology™ book series (MIMB, volume 65)


Genomic amplification with transcript sequencing (GAWTS) (1,2) 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 AMV reverse transcriptase (Fig. 1).
Fig. 1.

Schematic of GAWTS. GAWTS consists of the following three steps: (1) PCR, in which one or both oligonucleotides contain a phage promoter in addition to a sequence targeting the primer to the region to be amplified, (2) transcription with the phage promoter, and (3) dideoxy sequencing of the transcript with reverse transcriptase that is primed with a nested (internal) oligonucleotide. Reprinted with permission of Academic Press from Sommer et al. (2).


Xylene Cyanol Reverse Transcriptase Buffer Formamide Solution Shadow Band Dideoxynucleotide Sequencing 
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Copyright information

© Humana Press Inc., Totowa, NJ 1996

Authors and Affiliations

  • Tammy Lind
    • 1
  • Erik C. Thorland
    • 1
  • Steve S. Sommer
    • 1
  1. 1.Department of Biochemistry and Molecular BiologyMayo Clinic/FoundationRochester

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