An automated gridding procedure for the inoculation of yeast and bacterial clones in high-density arrays has been developed. A 96-pin inoculating tool compatible with the standard microtiter plate format and an eight-position tablet have been designed to fit the Biomek 1000 programmable robotic workstation (Beckman Instruments). The system is used to inoculate six copies of 80 x 120-mm filters representing a total of ~20,000 individual clones in approximately 3 h. High-density arrays of yeast artificial chromosome (YAC) and cosmid clones have been used for rapid large-scale hybridization screens of ordered libraries. In addition, an improved PCR library screening strategy has been developed using strips cut from the high-density arrays to prepare row and column DNA pools for PCR analysis. This strategy eliminates the final hybridization step and allows identification of a single clone by PCR in 2 days. The development of automated gridding technology will have a significant impact on the establishment of fully versatile screening of ordered library resources for genomic studies.
- DNA library
- molecular cloning
- polymerase chain reaction
- yeast artificial chromosome
Bentley, D. R., Todd, C., Collins, J., Holland, J., Dunham, I., Hassock, S., Bankier, A., & Giannelli, F. (1992). The development and application of automated gridding for efficient screening of yeast and bacterial ordered libraries. Genomics, 12(3), 534-541. https://doi.org/10.1016/0888-7543(92)90445-X