Heather Spillane, Stephen Milligan, Lynne Helton, Nibedita Mahanti, Kathy Kuebler, Dorothy Martus, Donald Yet, Paul Donald and Charles Barna
Michigan State Police DNA Laboratory, East Lansing, MI

× Ø × Ø × Ø × Ø × Ø × Ø × Ø × Ø × Ø × Ø × Ø × Ø × Ø × Ø × Ø

Chemiluminescent detection of forensic RFLP casework was implemented in our laboratory following an extensive validation study (presented at the 1995 Promega Symposium). Chemiluminescent detection was seen as a means of both reducing film exposure times and reducing unnecessary exposure to radioactive labeled probes. The transition from ³²P detection to alkaline phosphatase labeled oligonucleotide probes, Lumiphos Plus^tm substrate and Biodyne A^tm membranes has been successfully navigated, with relatively few problems. Improved sensitivity and reduced background was achieved, in the high volume casework environment, by making subtle adjustments to the prehybridization, hybridization and stringency protocols originally presented in the validation.

Casework membranes are routinely probed with D1S7, D2S44, D4S139, D5S110, D10S28, and D17S79. Membranes are ramped overnight and exposed for 30 minutes, on average. Optimum sensitivity is achieved with 200-400ng of DNA. Typical casework examples as well as the current protocol will be presented.