The deoxytetradecamer d(AAAACGACGGCCAG) has been shown to be an excellent universal primer for sequence determination of DNA cloned into the bacteriophage M13 mp7, mp8, and mp9 series. This new primer offers several advantages over others currently available and it has been used to define the cloning of Hinf I fragments of bacteriophage S13 DNA into the Eco RI site of M13 mp7, utilizing the homologous complementary base pairing of the two restriction sites. Of the four possible sequence derivatives of the Hinf I GANTC recognition site, only those corresponding to GAATC and GATTC have been found at cloning sites in chimeras.
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