Assembly of High-Resolution Restriction Maps Based on Multiple Complete Digests of a Redundant Set of Overlapping Clones

Vertically integrated analysis of human DNA. Final technical report

“…However, the system is clearly works and one technician could support the mapping demands of a group sequencing approximately Mbp/yr, starting with a YAC contigs. Although the quantitative agreement between fragment sizes in the maps and those predicted by the sequence is not bad (Gillett et al, 1996), we are capable of obtaining better fragment sizing, as documented in Riles et al (1 993). The current experimental system has been in too much flux in recent months to make optimization of fragment sizing a feasible objective.…”

“…This error rate is still not satisfactory, but two points stand out: all errors were correctly recognized because they led to MCD mapping inconsistencies, and the final map was sufficiently accurate to provide a meaningful quality-control check on the DNA sequence. The latter point is discussed in detail in Gillett et al (1996). The prediction of a single extra EcoRI site in the sequence-derived map is the error level expected if random sequencing errors occurred in random sequence at a frequency of 0.12%, which is probably fairly typical of the error rates now present in DNA sequence that is produced carefully.…”

“…The Gillett et al (1996) manuscript describes the most rigorous test of MCD mapping yet carried out on real data. By employing two probes spaced roughly one cosmid-length apart, DNA was sampled for a 1 15-kbp segment of yeast chromosome I11 from a deep, whole-genome cosmid library .…”

“…Images are acquired on a Fluorlmager 575 (Molecular Dynamics, Sunnyvale, CA) and analyzed using custom-developed image-analysis s o h a r e . Map assembly is carried out by a custom-developed software package developed in collaboration with W, Gillett of the Washington University Department of Computer Science (Gillett et al 1996).…”

“…An extensive discussion of the relative merits of this strategy and various alternatives is provided in the Introduction to our manuscript (Gillett et al, 1996). MCD mapping is a direct extension of the single-complete-digest method employed to map the S .…”