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 pSacBII P1 Cloning Vector and Three Cosmid Cloning Vectors: pTCF, svPHEP, and LAWRIST16
Pan, H., Wang, S., Chissoe, S., Bodenteich, A., Iyer, K., Clifton, S., Crabtree, J., & Roe, B. (1994). The Complete Nucleotide Sequences of the pSacBII P1 Cloning Vector and Three Cosmid Cloning Vectors: pTCF, svPHEP, and LAWRIST16. Genetic Analysis, Techniques and Applications, 11(5-6), 181 - 186. http://www.sciencedirect.com/science/article/pii/1050386294900396?via%3Dihub