RFLP band size standards: NIST standard reference material 2390

Area of Science:

• Forensic Science
• Genetics
• Biotechnology

Background:

• The U.S. advisory board mandates annual confirmation of forensic DNA profiling systems.
• National Institute of Standards and Technology (NIST) Standard Reference Material (SRM) 2390 is crucial for this confirmation.
• Previous recertification of SRM 2390 occurred in 1992.

Purpose of the Study:

• To recertify NIST SRM 2390 for HaeIII restriction enzyme-based restriction fragment length polymorphism (RFLP) profiling.
• To establish quantitative band size values for specific genetic loci using SRM 2390.
• To provide methods for validating RFLP systems and control materials against NIST standards.

Main Methods:

• An among-laboratory study involving 20 forensic laboratories was conducted.
• Data from two different-source DNAs (K562 cell line and TAW male DNA) were analyzed.
• Quantitative band sizes and bivariate tolerance intervals were determined for six genetic loci (D1S7, D2S44, D4S139, D5S110, D1OS28, D17S79).

Main Results:

• Quantitative band size values were established for the specified genetic loci.
• Bivariate tolerance intervals were determined for the band size data.
• The study confirmed SRM 2390's suitability for HaeIII RFLP profiling.

Conclusions:

• NIST SRM 2390 is a suitable standard for HaeIII RFLP profiling systems.
• The recertification study provides updated quantitative data for forensic DNA analysis.
• Detailed methods are provided for validating RFLP measurement systems and tracing them to NIST SRM 2390.