Announcement of Population Data Allele frequency distribution for 21 autosomal STR loci in Bhutan Thirsa Kraaijenbrink a , George L. van Driem b , Karma Tshering of Gaselo ˆ b , Peter de Knijff a, * a Department of Human Genetics, Postzone S5-P, Leiden University Medical Centre, P.O. Box 9600, 2300 RC, Leiden, The Netherlands b Himalayan Languages Project, FBJ Kuiperinstituut, Leiden University, P.N. van Eyckhof 3, 2311 BV Leiden, The Netherlands Received 1 March 2006; received in revised form 5 April 2006; accepted 11 April 2006 Available online 24 May 2006 Abstract We studied the allele frequency distribution of 21 autosomal STR loci contained in the AmpFlSTR 1 Identifiler TM (Applied Biosystems), the Powerplex 1 16 (Promega) and the FFFL (Promega) multiplex PCR kits among 936 individuals from the Royal Kingdom of Bhutan. As such these are the first published autosomal DNA results from this country. # 2006 Elsevier Ireland Ltd. All rights reserved. Keywords: Bhutan; Identifiler; Powerplex 1 16; FFFL Population information: The Royal Kingdom of Bhutan lies just south of the highest peaks in the eastern section of the Himalayan mountain range, which runs from Pakistan in the west to north-eastern India (Assam) in the east. As such, the Himalayas are not only the highest land barrier on earth, they also form a distinct barrier between Tibeto-Burman, which is predominantly spoken north-east of the Himalayas, and Indo- European, which is spoken south of the Himalayas [1]. In Bhutan, nearly all people speak a language belonging to the Tibeto-Burman language family. There are many distinct isolated languages within Bhutan, many even not completely characterized. It has been suggested that in the past humans speaking a Tibeto-Burman language either have crossed the Himalayas, or have navigated around the highest parts of the Himalayas before settling in Bhutan. In order to be able to reconstruct possible migration routes connecting human populations North and South of the Himalayas, we collected blood samples from 761 males and 175 females from Bhutan, belonging to 17 different language groups defined by van Driem [1] (see Fig. 1). Here we describe the first results of this study in the form of the allele frequency distributions and summary statistics of 21 different autosomal STR loci from the combined group of 936 Bhutanese individuals. To our knowledge, this is the first genetic study on autosomal STR loci from the Royal Kingdom of Bhutan. DNA extraction: DNA was extracted from whole blood, using the Autopure LS 1 from Gentra Systems, according to the manufacturer’s specifications. PCR and genotyping: Identifiler TM , Powerplex 1 16 and FFFL 1 PCR reactions were performed according to the manufacturers’ specifications. PCR products were analysed using an ABI 3100 automated DNA sequencer and the Genemapper 1 ID software. Statistical analysis of data: Allele frequencies and heterozygosity values (observed and expected) were calculated using GenePop version 3.4 [2], exact Hardy–Weinberg p-values and PIC values were calculated using PowerMarker [3]. The Excel PowerStats spreadsheet [4] was used to calculate power of discrimination and power of exclusion. We used the exact test of population differentiation included in Arlequin Ver. 2000 [5] to compare differences in allele frequencies among different populations. Results: Allele frequencies and summary statistics are presented in Table 1. We identified a number of new alleles (i.e. not yet reported in the NIST Short Tandem Repeat DNA Internet DataBase [6]). These are shown underlined in Table 1. Using the PowerMarker software, markers D16S539, D8S1179, D21S11, FGA, TPOX and LPL were found to have significant HWE p-values (i.e. below the Bonferroni corrected p-value threshold of 0.0023). Similar results were obtained by using GenePop. Further, GenePop analysis showed significant heterozygote deficiency for FGA. Subsequently, in order to test for the presence of undetected null alleles as a cause for heterozygote deficiency, the data were analysed using the www.elsevier.com/locate/forsciint Forensic Science International 170 (2007) 68–72 * Corresponding author. Tel.: +31 71 526 9540; fax: +31 71 526 8278. E-mail address: knijff@lumc.nl (P. de Knijff). 0379-0738/$ – see front matter # 2006 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.forsciint.2006.04.006