The National Institute of Justice (NIJ), in conjunction with the National Forensic Science Technology Center (NFSTC), is sponsoring a series of workshops designed to help facilitate the transition of novel technologies into practice by operational crime laboratories. These Technology Transition Workshops, which highlight technologies developed under the NIJ’s forensic science research and development programs, are a critical component of NIJ’s research, development, testing, and evaluation (RDT&E) efforts.
The purpose of this workshop is to introduce the method of post-PCR cleanup to forensic DNA analysts, and to provide information regarding instrumentation currently available to automate this process. Post- PCR cleanup is a simple and inexpensive method that can be used to increase the low relative fluorescent units (RFUs) often observed in low level samples. The goal of the procedure is to increase the RFUs so they are above the laboratory’s calling threshold. Samples that previously resulted in data that could not be used to make any forensically significant conclusions can now be subjected to post-PCR cleanup, allowing more definitive conclusions to be formed.
With the expansion of the use of DNA to solve more property crimes and with some agencies choosing DNA over latent prints to determine who was holding an item, analysis of low level samples is becoming more and more common. This technology can significantly improve the results from these low level samples so statements that would formerly have been “inconclusive” can now be statements of “inclusion” or “match”. Validation requirements and National DNA Index System (NDIS) admissibility issues will be addressed. This workshop will include both lecture and laboratory components.
At the completion of the MinElute® Post-PCR Cleanup workshop the participant will acquire the knowledge to:
- Explain how the post-PCR cleanup system works
- Discuss the strengths and limitations of the system
- Perform the method of post-PCR cleanup manually
- Perform the method of post-PCR cleanup using the QIAcube® robot
- Understand the validation method needed to introduce the methodology into the laboratory system
- Understand the downstream effects of using the method for low level samples and the uploading of those resulting profiles into CODIS
- Explain other uses of the QIAcube® robot
The following materials are provided as additional resources to this Workshop:
L. Forster, et al., Direct comparison of post-28-cycle PCR purification and modified capillary electrophoresis methods with the 34-cycle “low copy number” (LCN) method for analysis of trace forensic DNA samples, Forensic Sci. Int. Gene. (2008), doi:10.1016/j.fsigen.2008.04.005 Article (PDF 616kB)
Kathleen A. Mayntz-Press,1,4 M.S.; Lynn M. Sims,2,4 B.S.; Ashley Hall,2,4 Ph.D.; and Jack Ballantyne,1,2,3,4 Ph.D., Y-STR Profiling in Extended Interval (≥3 days) Postcoital Cervicovaginal Samples, J Forensic Sci, March 2008, Vol. 53, No. 2, doi: 10.1111/j.1556-4029.2008.00672.x Article (PDF 944kB)
Pamela J. Smith,1,2 M.S. and Jack Ballantyne,1,3,4 Ph.D., Simplified Low-Copy-Number DNA Analysis by Post-PCR Purification, J Forensic Sci, July 2007, Vol. 52, No. 4, doi: 10.1111/j.1556-4029.2007.00470.x Article (PDF 612kB)
User-Developed Protocol: Forensic post-PCR purification protocol using the MinElute® PCR Purification Kit Handout (PDF 244kB)