The complete nucleotide sequence of the 16,009-bp SacBII Kan domain of the P1 pAD10-SacBII cloning vector and the sequences of three cosmid cloning vectors, pTCF (7941 bp), svPHEP (9201 bp), and LAWRIST16 (5194 bp) have been determined. A modified diatomaceous earth (Prep-A-Gene)-based procedure, which rapidly yields highly supercoiled double-stranded DNA from recombinant P1 and cosmid clones suitable for generating shotgun libraries, also has been developed. The isolated recombinant DNAs were physically sheared to generate 1- to 2-kb fragments that then were blunt-ended and subcloned into double-stranded pUC-based sequencing vectors. The double-stranded sequencing templates were isolated by an alkaline lysis method and subjected to Taq polymerase catalyzed fluorescent end-labeled primer cycle sequencing. After shotgun sequence assembly, contig gaps were closed and ambiguities were resolved via Sequenase catalyzed fluorescent dye-terminator sequencing.
The complete nucleotide sequences of the SacBII Kan Domain of the P1 pAD10-SacBII cloning vector and three cosmid cloning vectors
pTCF, svPHEP, and LAWRIST16
Pan, H., Wang, Y-P., Chissoe, S. L., Bodenteich, A., Wang, Z., Iyer, K., Clifton, S. W., Crabtree, J. S., & Roe, B. A. (1994). The complete nucleotide sequences of the SacBII Kan Domain of the P1 pAD10-SacBII cloning vector and three cosmid cloning vectors: pTCF, svPHEP, and LAWRIST16. Genetic Analysis, Techniques and Applications, 11(5-6), 181-186. https://doi.org/10.1016/1050-3862(94)90039-6
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