Human Y Chromosome Dna Profiling Standard Srm 2395

SRM 2395 was released in July 2003 for use with verifying results involving Y chromosome STR testing (see Chapter 9). SRM 2395 includes five male DNA

samples (50 ||L at 2ng/|lL) selected to exhibit a diverse set of alleles across 31 commonly used Y chromosome STR and 42 single nucleotide polymorphism (SNP) markers. A female DNA sample is also included to serve as a negative control for male-specific DNA tests. In addition to the typing results from all commercially available Y-STR kits, the five male samples in SRM 2395 have been sequenced at 22 Y-STR loci to confirm allele calls (Kline et al. 2003b).

QUALITY CONTROL FOR COMMERCIAL SOURCES OF MATERIALS

In the early days of STR typing, forensic laboratories put together their own PCR mixes, primer sets, and allelic ladders. This meant that variation existed in the materials used for various laboratories and sometimes in the interpretation of a sample's genotype. Laboratories often had to spend a significant amount of time preparing the allelic ladders and verifying that each lot of primer mix worked appropriately. Today, most forensic DNA laboratories use commercially available STR kits that provide a uniform set of materials and protocols for the community. Thus, the primary responsibility of performing quality control on the STR amplification reagents has fallen on the commercial manufacturers.

Production of STR kits by commercial manufacturers requires extensive quality control. A fluorescent dye is attached to one primer for each locus amplified by the multiplex STR kit. Each primer must be purified and combined in the correct amount in order to produce a balanced amplification. Variation in this primer mix production can affect locus-to-locus balance in the multiplex amplification. In addition, allelic ladders must also be produced on a large scale and be well characterized since they serve as the standard for performing the DNA typing experiments with unknown samples. As they may be needed in some situations, Certificates of Analysis are available upon request from STR kit manufacturers for court purposes. The certificate confirms that the specific combination of components that comprise a given kit lot number perform together to meet the stated performance (Applied Biosystems 1998).

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CHAPTER 17

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