| Comments |
Restriction digests of the clone give the following sizes (kb): SalI--uncut; HindIII--22, 11, 5.9, 4.5, 1.5, 1.0; EcoRI--20, 9.8, 7.1, 3.5, 2.5, 1.1, 0.9, 0.7; PstI--7.1, 4.2, 3.3, 2.7, 2.2, 1.9, 1.7, 1.3, 1.1, 1.0, 0.9 + smaller; BamHI--9.8, 8.7, 7.9, 7.1, 3.1, 2.5, 2.5, 1.1. 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. Constant fragment sizes are 3.4, 2.3, 2.1, 1.44, 1.29, 1.16, 1.03, 0.94, and 0.84 kb. The same locus is revealed with a EcoRI/HindIII digest. This locus is in equilibrium with that detected by p37-1-34; the compound locus has a PIC of 0.54. A 20 kb EcoRI fragment probe can be obtained by digesting cosmid DNA with EcoRI and cutting the largest (20 kb) fragment out of the agarose gel. This is single-copy and gives good results with standard hybridization and washing conditions. |
