This is a method for screening YAC libraries for specific DNA sequences. Transformants are patched onto selective medium plates and colonies lifted to nylon filters for further growth.
Cells are then lysed in place and the DNA is fixed to the filters.
DNA probes are hybridized to the filters to test for the presence of specific sequences. A secondary screen is performed to ensure a positive hybridization. A protocol for purifying yeast artificial chromosomes by gel electrophoresis and retrieving the resultant products. To subclone the large insert fragment of human DNA contained within a YAC into a bacteriophage lambda vector. To identify the YAC subclones containing both a human insert and a portion of either the left or right arm of the pYAC4 vector.