Restriction digests analyzed on agarose gels give the following sizes (kb): EcoRI--5.7, 1.2, 0.56, 0.28; Sau3A--1.7, 0.88, smaller fragments; HindIII: 7.4; PstI--5.0, 2.4; SalI--7.4. EcoRI cuts the insert twice giving fragments of 1.2, 0.54 and 0.28 kb. 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. This probe detects 3 different TaqI polymorphisms. Alpha (C alleles) is the same detected by p13-1-82 (ATCC 59004, 59005). Epsilon has more than 32 alleles. Phi (F alleles) has 2 common alleles. The combined heterozygosity for these is 98%. The highly polymorphic epsilon polymorphism includes a relatively frequent null allele. Can use the entire plasmid as probe with standard hybridization and washing conditions. Use a final wash of 0.1X SSC/0.1% SDS at 65C. p13-1-82 (ATCC 59004, 59005), p13-2-31 (ATCC 59014, 59015), and p13-1-25 (ATCC 59870, 59871) were subcloned from cosmid 1-13 (C1-13). |
