scholarly journals ANALISIS PENGARUH PAPARAN FISIK PADA SAMPEL GIGI TERHADAP HASIL KUANTIFIKASI DNA FORENSIK MENGGUNAKAN METODE KIT PURIFIKASI DNA KOMERSIAL

2021 ◽  
Vol 5 (2) ◽  
pp. 59-65
Author(s):  
Septi Arini ◽  
Arief Budi Witarto ◽  
Setia Betaria Aritonang
Keyword(s):  
Real Time ◽  
River Water ◽  
Real Time Pcr ◽  
Room Temperature ◽  
Sea Water ◽  
Water Immersion ◽  
Dna Quantification ◽  
Forensic Dna ◽  
Physical Exposure ◽  
Free Air

Physical exposure to biological samples has an enormous influence on the results of forensic DNA analysis. The lack of molecular research on the effect of physical exposure on dental samples is the reason for the need for further research. This study aims to determine the effect of physical exposure on dental samples on the results of forensic DNA quantification. The parameter used is the concentration value of isolated DNA obtained from real time PCR analysis. The use of real time PCR allows the detection and quantification of specific sequences of DNA samples at the same time to be analyzed. The dental samples used were obtained from different individuals. Teeth are used as identification media because teeth are the hardest part of the body and are chemically the most stable and most resistant to degradation and decomposition. The method in this study is to give three types of treatment to the tested samples in the form of sea water immersion, river water immersion and exposure to free air at room temperature with each treatment consisting of three test samples. All samples were extracted using a Commercial DNA purification KIT with a reagent in the form of a Qiagen KIT (QIAamp® DNA Investigator) then a quantification process was carried out to see the value of the DNA concentration of each sample using real time PCR. The results of DNA quantification of dental samples from each treatment showed that the highest sample concentration value was based on the average of each treatment, namely samples with treatments exposed to free air at room temperature with a concentration value of 1.34 ng/µl, followed by samples soaked using river water with a concentration value of 0.15 ng/µl, while the sample with the lowest concentration is shown by a sample treated with seawater immersion with a concentration value of 0.10 ng/µl. Physical exposure in the form of exposure to free air, exposure to river water and exposure to sea water on dental samples, gave a not too significant effect on the results of DNA quantification produced.

Real-time PCR detection of adenoviruses, polyomaviruses, and torque teno viruses in river water in Japan

2010 ◽  
Vol 44 (6) ◽  
pp. 1747-1752 ◽  
Author(s):  
Eiji Haramoto ◽  
Masaaki Kitajima ◽  
Hiroyuki Katayama ◽  
Shinichiro Ohgaki
Keyword(s):  
Real Time ◽  
River Water ◽  
Real Time Pcr ◽  
Pcr Detection

scholarly journals Multilaboratory Evaluation of Real-Time PCR Tests for Hepatitis B Virus DNA Quantification

2011 ◽  
Vol 49 (8) ◽  
pp. 2854-2858 ◽  
Author(s):  
A. M. Caliendo ◽  
A. Valsamakis ◽  
J. W. Bremer ◽  
A. Ferreira-Gonzalez ◽  
S. Granger ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Real Time ◽  
Real Time Pcr ◽  
Dna Quantification ◽  
Hepatitis B Virus Dna ◽  
B Virus

scholarly journals CMV DNA quantification in plasma using an automated real-time PCR assay

2006 ◽  
Vol 36 ◽  
pp. S6-S7
Author(s):  
S. Yerly ◽  
L. Kaiser ◽  
C. Van Delden ◽  
S. Schaffer ◽  
S. Thamm ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Dna Quantification ◽  
Pcr Assay

scholarly journals Stabilization of circulating thyroglobulin mRNA transcripts in patients treated for differentiated thyroid carcinoma

2016 ◽  
Vol 54 (5) ◽  
pp. 558-566 ◽  
Author(s):  
Søren Feddersen ◽  
Lars Bastholt ◽  
Susanne M Pedersen
Keyword(s):  
Thyroid Carcinoma ◽  
Real Time ◽  
Real Time Pcr ◽  
Room Temperature ◽  
Differentiated Thyroid Carcinoma ◽  
Mrna Levels ◽  
Serum Thyroglobulin ◽  
Tempus Tubes ◽  
Antithyroglobulin Antibodies ◽  
Previously Treated

Background The clinical utility of serum thyroglobulin in the follow-up of patients with differentiated thyroid carcinoma may be compromised by the presence of endogenous antithyroglobulin antibodies. To prevent interference by antithyroglobulin antibodies several groups have developed real-time PCR-based assays for quantification of blood thyroglobulin mRNA levels. For accurate quantification of thyroglobulin mRNA in blood preanalytical factors must be recognized and controlled. In this study, we evaluate the effect of different blood RNA stabilizing systems – the Tempus Blood RNA system and the PAXgene Blood RNA system – and storage time on RNA yield and quality, and thyroglobulin mRNA stability. Methods Blood samples from 11 patients previously treated for differentiated thyroid carcinoma were collected in K2-EDTA, Tempus and PAXgene tubes and maintained at room temperature. RNA was isolated following storage for 0 and 72 h, and RNA yield, integrity and purity was determined. Thyroglobulin, GAPDH and ACTB mRNA levels were quantified by semi-quantitative real-time PCR. Results The RNA yield was significantly higher for blood collected in Tempus tubes compared with PAXgene tubes following storage for 72 h at room temperature ( P = 0.0011). High-quality RNA could be extracted from blood collected in PAXgene and Tempus tubes. Blood collected in K2-EDTA tubes, but not in PAXgene and Tempus tubes, showed significant changes in thyroglobulin mRNA levels following storage for 72 h at room temperature ( P = 0.0263). Conclusions Stabilization of blood in PAXgene and Tempus tubes enables storage at room temperature for up to 72 h, without compromising thyroglobulin mRNA levels.

Quantitative detection of human enteric adenoviruses in river water by microfluidic digital polymerase chain reaction

2014 ◽  
Vol 70 (3) ◽  
pp. 555-560 ◽  
Author(s):  
Naohiro Kishida ◽  
Naohiro Noda ◽  
Eiji Haramoto ◽  
Mamoru Kawaharasaki ◽  
Michihiro Akiba ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
River Water ◽  
Water Samples ◽  
Real Time Pcr ◽  
Quantitative Detection ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction ◽  
Enteric Adenoviruses

We describe an assay for simple and accurate quantification of human enteric adenoviruses (EAdVs) in water samples using a recently developed quantification method named microfluidic digital polymerase chain reaction (dPCR). The assay is based on automatic distribution of reaction mixture into a large number of nanolitre-volume reaction chambers and absolute copy number quantification from the number of chambers containing amplification products on the basis of Poisson statistics. This assay allows absolute quantification of target genes without the use of standard DNA. Concentrations of EAdVs in Japanese river water samples were successfully quantified by the developed dPCR assay. The EAdVs were detected in seven of the 10 samples (1 L each), and the concentration ranged from 420 to 2,700 copies/L. The quantified values closely resemble those by most probable number (MPN)-PCR and real-time PCR when standard DNA was validated by dPCR whereas they varied substantially when the standard was not validated. Accuracy and sensitivity of the dPCR was higher than those of real-time PCR and MPN-PCR. To our knowledge, this is the first study that has successfully quantified enteric viruses in river water using dPCR. This method will contribute to better understanding of existence of viruses in water.

scholarly journals Methods for DNA quantification yield similar relative but different absolute values

2019 ◽  
pp. 25-31
Author(s):  
O. P. Balanovsky ◽  
ZhA Kagazezheva ◽  
M. V. Olkova
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Genomic Dna ◽  
Correlation Coefficients ◽  
Dna Quantification ◽  
Sample Storage ◽  
Excellent Correlation ◽  
The Real ◽  
Absolute Concentrations

DNA quantification is a routine yet important procedure that determines the efficacy of long-term sample storage and further manipulations with the sample. There are a few well-established methods for measuring DNA concentrations. However, it still not fully clear how concordant their results are. The aim of this work was to measure DNA concentrations in a set of samples using different quantification methods and to compare the obtained values. In 2 independent experiments, a total of 100 genomic DNA samples were analyzed using 3 different DNA quantification methods, including spectrophotometry (NanoDrop), fluorometry (Qubit) and real-time PCR (Quantifiler). The obtained relative concentrations demonstrated an excellent correlation (the correlation coefficients were as high as 0.98 to 0.99). However, the absolute concentrations showed a considerable variation and even a twofold difference. Spectrophotometry yielded the highest concentrations, whereas fluorometry yielded the lowest. The real-time PCR results were intermediate. The differences were more pronounced for the samples with low DNA concentrations. We recommend that such differences should be accounted for when estimating DNA concentrations using an arsenal of different quantification methods.

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