Direct PCR amplification of the HVSI region in mitochondrial DNA from buccal cell swabs
2012
Autori
Kovačević Grujičić, NatašaDavidović, Slobodan
Takić, Dijana
Mojsin, Marija
Stevanović, Milena
Članak u časopisu (Objavljena verzija)
Metapodaci
Prikaz svih podataka o dokumentuApstrakt
Amplification of human mitochondrial DNA (mtDNA) has been widely used in population genetics, human evolutionary and molecular anthropology studies. mtDNA hypervariable segments I and II (HVSI and HVSII) were shown to be a suitable tool in genetic analyses due to the unique properties of mtDNA, such as the lack of recombination, maternal mode of inheritance, rapid evolutionary rate and high population-specific polymorphisms. Here we present a rapid and low-cost method for direct PCR amplification of a 330 bp fragment of HVSI from buccal cell samples. Avoiding the DNA isolation step makes this method appropriate for the analysis of a large number of samples in a short period of time. Since the transportation of samples and fieldwork conditions can affect the quality of samples and subsequent DNA analysis, we tested the effects of long-term storage of buccal cell swabs on the suitability of such samples for direct PCR amplification. We efficiently amplified a 330 bp fragment of HVSI even... after the long-term storage of buccal cells at room temperature, +4°C or at -20°C, for up to eight months. All examined PCR products were successfully sequenced, regardless of sample storage time and conditions. Our results suggest that the direct PCR amplification of the HVSI region from buccal cells is a method well suited for large-scale mtDNA population studies.
Ključne reči:
mtDNA / HVSI / direct PCR / buccal swabIzvor:
Archives of Biological Sciences, 2012, 64, 3, 851-858Izdavač:
- Srpsko biološko društvo, Beograd, i dr.
Finansiranje / projekti:
- Etnogeneza Srba u srednjem veku: Uporedna analiza istorijsko-kulturnog nasleđa, genetičkog materijala i predmeta materijalne kulture sa aspekta analitičke hemije (RS-MESTD-Integrated and Interdisciplinary Research (IIR or III)-47025)
DOI: 10.2298/ABS1203851G
ISSN: 0354-4664
WoS: 000308511300004
Scopus: 2-s2.0-84867884460
Institucija/grupa
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Kovačević Grujičić, Nataša AU - Davidović, Slobodan AU - Takić, Dijana AU - Mojsin, Marija AU - Stevanović, Milena PY - 2012 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/578 AB - Amplification of human mitochondrial DNA (mtDNA) has been widely used in population genetics, human evolutionary and molecular anthropology studies. mtDNA hypervariable segments I and II (HVSI and HVSII) were shown to be a suitable tool in genetic analyses due to the unique properties of mtDNA, such as the lack of recombination, maternal mode of inheritance, rapid evolutionary rate and high population-specific polymorphisms. Here we present a rapid and low-cost method for direct PCR amplification of a 330 bp fragment of HVSI from buccal cell samples. Avoiding the DNA isolation step makes this method appropriate for the analysis of a large number of samples in a short period of time. Since the transportation of samples and fieldwork conditions can affect the quality of samples and subsequent DNA analysis, we tested the effects of long-term storage of buccal cell swabs on the suitability of such samples for direct PCR amplification. We efficiently amplified a 330 bp fragment of HVSI even after the long-term storage of buccal cells at room temperature, +4°C or at -20°C, for up to eight months. All examined PCR products were successfully sequenced, regardless of sample storage time and conditions. Our results suggest that the direct PCR amplification of the HVSI region from buccal cells is a method well suited for large-scale mtDNA population studies. PB - Srpsko biološko društvo, Beograd, i dr. T2 - Archives of Biological Sciences T1 - Direct PCR amplification of the HVSI region in mitochondrial DNA from buccal cell swabs EP - 858 IS - 3 SP - 851 VL - 64 DO - 10.2298/ABS1203851G ER -
@article{ author = "Kovačević Grujičić, Nataša and Davidović, Slobodan and Takić, Dijana and Mojsin, Marija and Stevanović, Milena", year = "2012", abstract = "Amplification of human mitochondrial DNA (mtDNA) has been widely used in population genetics, human evolutionary and molecular anthropology studies. mtDNA hypervariable segments I and II (HVSI and HVSII) were shown to be a suitable tool in genetic analyses due to the unique properties of mtDNA, such as the lack of recombination, maternal mode of inheritance, rapid evolutionary rate and high population-specific polymorphisms. Here we present a rapid and low-cost method for direct PCR amplification of a 330 bp fragment of HVSI from buccal cell samples. Avoiding the DNA isolation step makes this method appropriate for the analysis of a large number of samples in a short period of time. Since the transportation of samples and fieldwork conditions can affect the quality of samples and subsequent DNA analysis, we tested the effects of long-term storage of buccal cell swabs on the suitability of such samples for direct PCR amplification. We efficiently amplified a 330 bp fragment of HVSI even after the long-term storage of buccal cells at room temperature, +4°C or at -20°C, for up to eight months. All examined PCR products were successfully sequenced, regardless of sample storage time and conditions. Our results suggest that the direct PCR amplification of the HVSI region from buccal cells is a method well suited for large-scale mtDNA population studies.", publisher = "Srpsko biološko društvo, Beograd, i dr.", journal = "Archives of Biological Sciences", title = "Direct PCR amplification of the HVSI region in mitochondrial DNA from buccal cell swabs", pages = "858-851", number = "3", volume = "64", doi = "10.2298/ABS1203851G" }
Kovačević Grujičić, N., Davidović, S., Takić, D., Mojsin, M.,& Stevanović, M.. (2012). Direct PCR amplification of the HVSI region in mitochondrial DNA from buccal cell swabs. in Archives of Biological Sciences Srpsko biološko društvo, Beograd, i dr.., 64(3), 851-858. https://doi.org/10.2298/ABS1203851G
Kovačević Grujičić N, Davidović S, Takić D, Mojsin M, Stevanović M. Direct PCR amplification of the HVSI region in mitochondrial DNA from buccal cell swabs. in Archives of Biological Sciences. 2012;64(3):851-858. doi:10.2298/ABS1203851G .
Kovačević Grujičić, Nataša, Davidović, Slobodan, Takić, Dijana, Mojsin, Marija, Stevanović, Milena, "Direct PCR amplification of the HVSI region in mitochondrial DNA from buccal cell swabs" in Archives of Biological Sciences, 64, no. 3 (2012):851-858, https://doi.org/10.2298/ABS1203851G . .