Restriction digests of the clone give the following sizes (kb): EcoRI/BglII--4.9, 2.45, 0.67, 0.48; BamHI--10.0; BglII--10.0; EcoRI--5.3, 2.65, 1.25; HindIII--10.0. The GM1416 cell line was incorrectly described as 49,XXXXY. 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. The distance between the EcoRI RFLP (detected by F.53, ATCC 61108) and the MspI RFLP (detected by H5.52, ATCC 61106) is 48 kb. For detecting the RFLP, the complete insert should be used to get good signal. This probe was derived from the cosmid C5.5. C5.5 was isolated from a library constructed by digesting the vector C2RB with BamHI and SmaI and inserting size-selected fragments from GM1416 partially digested with MboI. This probe was derived from C5.5 by digesting with BglII and religating. This is from the more telomeric end of C5.5. The 3.6 kb DNA fragment in this clone contains an internal EcoRI site. An EcoRI/BglII digest of the clone gives 2 unique fragments from the insert: a 2.4 kb EcoRI fragment and a 1.2 kb EcoRI/BglII fragment. |
