Restriction digests of the clone give the following sizes (kb): PstI/SalI--5.6, 3.8; AvaI--3.5, 3.2, 1.44, 0.82 + smaller; EcoRI--9.4; BglI--7.55, 1.85; HindIII--9.4. The insert contains the following restriction sites (in approximate kb from the PstI site): NotI--0.44; BssHII--0.96. 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 is approximately 15 kb from DR47. Detects genomic restriction fragments of the following sizes (complete digest, kb): NotI--450; EagI--260; BsshII--160; SacII--160. The ClaI and the BamHI sites of the polylinker are absent as a result of the probe construction. All other sites of the polylinker are retained. This was subcloned from the end of the cosmid c26 (Cos 26) by digestion with PstI and religation of the vector/insert construct at low DNA concentration. |
