NUCLEAR DNA QUANTIFICATION OF BONE SAMPLES FROM DIFFERENT SOURCES: A COMPARISON BETWEEN REAL-TIME PCR, AGAROSE GEL ELECTROPHORESIS AND SPECTROPHOTOMETRY (CROSBI ID 530997)
Prilog sa skupa u zborniku | sažetak izlaganja sa skupa | međunarodna recenzija
Podaci o odgovornosti
Sutlović, Davorka ; Gugić, Dijana ; Definis-Gojanović, Marija ; Anđelinović, Šimun
engleski
NUCLEAR DNA QUANTIFICATION OF BONE SAMPLES FROM DIFFERENT SOURCES: A COMPARISON BETWEEN REAL-TIME PCR, AGAROSE GEL ELECTROPHORESIS AND SPECTROPHOTOMETRY
To comparate analytical methods to evaluate nuclear DNA concentrations of forensic and ancient bone samples. Three different methods were used: agarose gel electrophoresis, spectrophotometry and Quantitative Real-time PCR. QRT-PCR offers several advantages with respect to other compared methods. Twenty four DNA extracts from bone samples: eight DNA extracts from ancient bone samples (3 DNA samples retrieved from 10-50 years old bones and 5 DNA samples obtained from about 1300 years old bones recovered from an old church burrial sight), eight DNA were extraced from mass graves bones and eight were extracted from fresh bones. The quantification assay was performed in total volume of 25  L containing 2  L of DNA extract, Quantifiler human primer mix and Quantifiler PCR reaction mix with thermal cycling conditions according to the manufacture’ s protocols In the following trials total DNA was evaluated by gel electrophoresis (1.5% agarose) and ethidium bromide staining by UV iluminations (312 nm) of the gel . Absorbancies at 260 and 280 nm were measured by spectrophotometer. Highly sensitive methods for quantification of human DNA retrieved from forensic and ancient DNA samples are important because they ensure the consistency of low copy number DNA typing. RT-PCR offers several advantages with respect to other compared methods. In samples which contain high amounts of DNA all of the tested methods gave positive results with good agreement. All samples that possibly contain small amount of DNA or inhibitors therefore need to be quantitated by QRT-PCR as method of choice.
analitical methods; human DNA; quantification; bone samples
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Podaci o prilogu
2007.
objavljeno
Podaci o matičnoj publikaciji
Primorac, Dragan
Zagreb: Studio Hrg
Podaci o skupu
5th ISABS Conference in Forensic genetics and Molecular Anthropology
poster
03.09.2007-07.09.2007
Split, Hrvatska