The insert does not contain EcoRI, KpnI, SacI or BamHI sites. The SacI and XhoI sites from the multiple cloning site of the original lambda vector GEM-11 were not removed and are there. The insert contains a HindIII site 1.3 kb from the EcoRI end and another HindIII site 0.1 kb from the SacI end. Restriction digests of the clone give the following sizes (kb): EcoRI/SacI--3.0, 2.0; HindIII--4.4, 0.6; PstI--uncut; BamHI--5.0; XhoI--3.0, 2.0. Detects a genomic SacI fragment of 18 kb. Requires preassociation with sheared human DNA prior to hybridization for optimal results. Has been used successfully to detect transcripts in Northern blots. |
