Research Facility 

 Narratives: LLNL 



Instrumentation and Automation. The group responsible for this task develops 

 instrumentation that will facilitate the handling of DNA and cloned materials so that 

 human involvement in highly repetitive tasks can be minimized. 



High-Resolution Imaging. In this project, two novel approaches to DNA sequencing 

 are being developed using scanning tunneling microscopy (STM) and high-resolution 

 imaging with X-ray lasers. 



The research groups involved in these component tasks are highly interactive. Individ- 

 ual staff members often have responsibilities in more than one group. Collaborating 

 with other research groups throughout the world. Livennore coordinates its research 

 activities with others who are either involved directly in the human genome effort or 

 who have mutual scientific interests. 



Accomplishments 



In the past two years, Livermore has made excellent progress in the construction of 

 chromosome-specific libraries, in the development and application of new biochemical 

 and mathematical approaches for constructing ordered-clone maps, in the automation of 

 fingerprinting chemistries, and in high-resolution imaging of DNA. 



As part of the National Laboratory Gene Library Project, small insert libraries of each 

 human chromosome were constructed using material purified by flow sorting. These 

 libraries have been deposited with the American Type Culture Collection for worldwide 

 distribution. Construction of large-insert partial-digest libraries in both lambda and 

 cosmid vectors has begun: currently, five such libraries are being characterized for 

 chromosomes 11, 19, 21, 22, and Y. 



In the area of resource development, new or modified existing vectors have been 

 constructed to: 



• facilitate cloning small amounts of DNA in cosmids, 



• clone Not I linking probes in lambda and plasmids, and 



• clone large fragments of DNA as YACs, 



Several of the cosmid vectors have been transferred to the U.S. commercial sector. They 

 were used to construct cosmid libraries of flow-sorted chromosomes. The plasmid and 

 lambda vectors were used to create a small Not I linking library of chromosome 19. A 

 library of chromosome 19 in YACs is currently being expanded. A probe repository for 

 chromosome 19 has been established, and the collection is growing: these probes have 

 been used to screen Livermore's chromosome-19 libraries. Individual cosmid clones 

 and filters containing DNA from individual clones of chromosome 19 are being 

 distributed to the scientific community as part of a collaborative effort to map this 

 chromosome. 



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