Restriction digests of the clone give the following sizes (kb): EcoRI--4.8; EcoRI/HindIII--3.3, 1.5; HindIII--4.8; AccI--uncut; BamHI--4.8. For plasmid amplification, use a lacIq host. This was constructed with EcoRI site on the T7 (5') side and a destroyed PstI site on the T3 (3') side. The insert contains internal BglII, BamHI, and HincII sites. Codominant X-linked segregation of the HindIII polymorphism was observed in 4 families. ATCC 57666 has been converted to meet the requirements of the Budapest Treaty for patent deposits. A compliance form is not needed for ATCC 57666, but is for ATCC 57667. This insert includes probes 1 and 2a. It detects genomic HindIII fragments of (3.2, 3.25), 4.2, 8.5, 3.1, 8.0, 4.6, 7.5, and 10.5 kb. This probe encompasses the DXS142 (pERT84) 3.2 kb fragment, the J-MD 4.6 kb fragment, the DXS206 (XJ1) 7.5 kb fragment, and the DXS164 (pERT87-28) 10.5 kb fragment. Of the DMD cDNA clones, 1-2a includes nucleotides 1-1538; 2b-3: 1455 to approximately 2600; 4-5a: 2600-4550; 5b-7: 4400-6900; 8: 6900-7800; 9-14: 7800-13900 (in base pairs from the 5' end of the cDNA). A 0.6% agarose gels is required to resolve the bands of the HindIII RFLP. Absence of the 7.5 kb allele may be misinterpreted due to poor resolution of the 8.3 kb allele from the constant 8.0 and 8.5 kb bands on 0.8% agarose gels. Enzyme(s) not detecting polymorphism: AluI, BglII, EcoRI, EcoRV, HaeIII, HinfI, MspI, MvaI, PstI, TaqI. |
