Development and validation of a multiplexed Y-chromosome STR genotyping system, Y-PLEX 6, for forensic casework.
A Y-chromosome multiplex polymerase chain reaction (PCR) amplification kit, known as Y-PLEX 6, has been developed for use in human identification. The Y-PLEX 6 kit enables simultaneous amplification of six polymorphic short tandem repeat (STR) loci located on the non-recombinant region of the human Y-chromosome. These loci are: DYS393, DYS19, DYS38911, DYS390, DYS391, and DYS385. Our studies show that as little as 0.2 ng of template DNA can be used for analysis. The specificity of the amplification reaction enabled analysis of male DNA in a male:female DNA mixture at a ratio of 1:125. Among the six Y-STR loci, the maximum mean stutter percentage was 11.9 for allele at DYS38911 locus. Attempts at amplification of DNA from various animal sources revealed that the Y-PLEX 6 primers are human specific. Details of the development of the kit, generation and description of the allelic ladders, and validation of the multiplex PCR are presented. In addition, Y-STR allele and haplotype frequencies in three populations have been investigated. The data indicate that results obtained using the Y-PLEX 6 kit are robust, sensitive, and reliable and can be used in human forensic and male lineage identification cases.
Sinha
Sudhir K
SK
ReliaGene Technologies, Inc. 5525 Mounes St. Suite 101, New Orleans, LA 70123, USA. sinha@reliagene.com
Budowle
Bruce
B
Arcot
Santosh S
SS
Richey
Stephen L
SL
Chakrabor
Ranajit
R
Jones
Mary D
MD
Wojtkiewicz
Patrick W
PW
Schoenbauer
Dolores A
DA
Gross
Ann Marie
AM
Sinha
Siddhartha K
SK
Shewale
Jaiprakash G
JG
D000818:Animals; D016172:DNA Fingerprinting / Q000379:methods*; D005260:Female; D005554:Forensic Medicine / Q000379:methods; D005828:Genetics, Population; D005838:Genotype; D006239:Haplotypes; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D044469:Racial Groups / Q000235:genetics; D012680:Sensitivity and Specificity; D019849:Sex Determination Processes; D020080:Tandem Repeat Sequences; D014998:Y Chromosome