Restriction digests of the clone give the following sizes (kb): EcoRI/HindIII--2.4 doublet, 1.2, 1.0; EcoRI--5.1, 2.4 doublet; HindIII--3.8, 2.4, 1.2; consistent with deposited DNA. The structure of this plasmid is not consistent with the published description of pHHH202. The hybridization pattern of this probe with human DNA was not checked by the ATCC. This plasmid shows some instability. We recommend growing on LB agar with ampicillin, and picking colonies to select the correct clone. Digests also seem to indicate there is a high percentage of vectors without insert present. Linked to von Recklinghausen neurofibromatosis (NF1) (theta = 0.001, LOD score = 12.7). The probe detects genomic HindIII fragments of 2.4 and 1.6 kb and does not cross-hybridize to rat or mouse sequences. Codominant segregation demonstrated in 49 3-generation families for the RsaI RFLP. This probe was pre-associated with excess human DNA before hybridization. Otherwise, RFLPs were observed under normal conditions of hybridization and washing. Enzyme(s) not detecting polymorphism: BamHI, BglII, EcoRI, HindIII, MspI, PstI, PvuII, TaqI. |
