HindIII/EcoRI digests give 3.0 kb (vector) and 1.7, 0.7, 0.4 kb (insert). The 1.7 kb fragment can be used as a probe. Sau3A digests give 2.0 kb (from insert, use as probe) + other smaller fragments. Restriction digests of the clone give the following sizes (kb): BglI--4.7, 2.0; PvuII--6.4; HindIII--6.4; EcoRI--5.08, 1.1, 0.7; HindIII/EcoRI--3.3, 2.2, 1.0, 0.7. Long gel runs are required to resolve RFLP alleles. Depositor suggests 1% gels, 20 hrs, ca 2 V/cm. This locus is in linkage equilibrium with that seen by the 20 kb EcoRI fragment probe of cosmid 1-37. The compound locus has a PIC of 0.54. Hybridizations are done in 50% formamide, etc. at 50C; wash at 0.1X SSC /0.1% SDS, 65-68C. Probe hybridizes weakly to repeat sequences in TaqI fragments larger than 7 kb. This was derived from cosmid 1-37 (ATCC 59010). The cosmid was digested with Sau3A1 and a 2.8 kb fragment was used to construct this clone. |
