| Insert |
DNA: genomic Insert lengths(kb): 3.200000047683716 Gene product: DNA Segment, single copy [D2S3] Alleles: C4, J10, A1, B1, B2, C1, C2, C3, D2, G4, J1, J3, J4, J5, J6, J7, J8, K2, A2, D1, E1, E2, G1, G2, G3, J2, J9, K1 |
| Comments |
Restriction digests of the clone give the following sizes (kb): EcoRI--5.4, 3.2; HindIII--8.6; EcoRI/HindIII--5.4, 3.2; PstI--6.0, 1.8, 0.88. IMPORTANT: To prevent amplification of a rearranged and/or deleted cosmid, we recommend streaking on LB + amp plates at 30C and picking small colonies for liquid culture. p5-1-25 is a single copy subclone from cosmid 1-5. The most efficient detection of RFLPs at the D2S3 locus is by use of the cosmid 1-5 subclones p5-1-25 (ATCC 59684) and p5-2-96 (ATCC 59000). Use of p5-1-30 (ATCC 59006) and p5-1-32 (ATCC 59002) probably adds little to the PIC. Subclone 5-1-32 detects the PstI insert/deletion RFLP with TaqI but bands are larger and difficult to resolve. Long runs on 20 cm long 1% agarose gels, such that all fragments less than about 1.5 kb have run off the end, are required to obtain adequate resolution. |
| References |
Litt M. A single-copy subclone, p1-25, from cosmid 1-5, defines a highly polymorphic RFLP on 2q35->37 [HGM8 no. D2S3]. Nucleic Acids Res. 14: 4378, 1986. PubMed: 3714483
Litt M, et al. A highly polymorphic locus in human DNA revealed by probes from cosmid 1-5 maps to chromosome 2q35-37. Am. J. Hum. Genet. 38: 288-296, 1986. PubMed: 3006480
Lathrop GM, et al. Three genetic linkage groups on chromosome 2. Cytogenet. Cell Genet. 46: 644, 1987.
Asher JH Jr., et al. Waardenburg syndrome (WS): the analysis of a single family with a WS1 mutation showing linkage to RFLP markers on human chromosome 2q. Am. J. Hum. Genet. 48: 43-52, 1991. PubMed: 1670751
Litt M, White RL. A highly polymorphic locus in human DNA revealed by cosmid-derived probes. Proc. Natl. Acad. Sci. USA 82: 6206-6210, 1985. PubMed: 2994065
Michael Litt, personal communication
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