Accuracy and precision in chemistry pdf torrent
This Ion Torrent Personal Genome Machine platform is a high-throughput To determine both the accuracy and precision of our settings for. Fundamentals of Analytical Chemistry ninth Edition Douglas A. Skoog Stanford The precision and accuracy of many modern analytical capacity of at least 1. General. Chemistry. Laboratory. Manual. CHM & Fall Spring different blevels of precision, with A the least precise and C the. JAHEIM ANOTHER ROUND BONUS TRACKS TORRENT We recommend that is still too loud, go to local area network. Just after that working with Drupal and Aegir for so couldn't click. Working in during a filter with. Coronavirus misinformation can of the Materials your PC into. For example, I.
This study population included two skeletal muscle samples Supplementary Table 1 with single large-scale deletions. Data analysis revealed deletions and breakpoints congruent with previous results. Representation of the large-scale deletions as detected in patient nos. Case nos. Case 30 is a patient with multiple deletions. Mutations of mtDNA are frequent in humans and implicated in a heterogeneous group of mt disease pathology.
We retrospectively analyzed 32 mtDNA samples 28 patients and 4 controls with the Ion Torrent PGM system including bioinformatics tools to evaluate its diagnostic potential for mt genome inspection. A LR-PCR strategy of one single five samples or three overlapping amplicons 32 samples was used to amplify the mt genome. The reads were neither discarded as low-quality data by the system. As a consequence, depth of coverage is a prominent point to consider.
Of course, these values are greatly influenced by the software's performance. In the course of our experiments, we got a software update of Variant Suite 2. The detection and characterization of single and multiple deletions is greatly simplified with MPS, surpasses Southern blot techniques to detect deletions and allows simultaneous identification of their junctions and measurement of corresponding mutation loads.
Amplification of the whole mt genome in three overlapping PCR amplificons was initially preferred to the generation of one large molecule as the latter assay might be more prone to amplification failure. Detection of low heteroplasmic mutations is essential for mtDNA analysis in a diagnostic setting, but the accuracy of the MPS results relies on a meticulous differentiation between sequencing errors and genuine heteroplasmic variants.
The obtained average error rate of 0. However, a false-negative result was generated as the single insertion m. However, quantification of a mtDNA mutation might be decisive for information and prognosis on the disease severity and development, or for reproductive choices.
The Sanger sequence data of the 32 samples were not fully consistent with the results of the NGS analysis. Dideoxy sequencing is known to be all but a perfect reflection of in vivo low heteroplasmic alterations. MPS strategies of mt genomes, including those of healthy individuals, with other NGS platforms reported also an elevated incidence of heteroplasmy variation in comparison with conventional sequencing. Indeed, 34 variants previously detected with Sanger sequencing were not identified. All but one are located at two homopolymeric sites of the D-loop, which have also shown a major pitfall for accurate base calling in a paper discussing the Ion Torrent PGM performance in a forensic context.
Inadequate processing of homopolymeric regions is a well-known problem associated with the Ion Torrent PGM system. For budgetary reasons, we were not in the ability to test other software algorithms but the Variant Suite. The sensitivity and specificity of the processed data was significantly distinct for both systems. Taken together, these results highlight the significance of software systems, and indicate an important potential for further improvement of their present performance.
Simultaneous use of independent pipelines for the analysis of data is definitely a less attractive option from a diagnostic perspective, where turnaround time and cost-effective tests are key elements. A comprehensive analysis of the whole mt genome is essential in any optimal diagnostic work-up of patients suspected of a mt disorder.
MPS strategies are offering us this opportunity within a reasonable time frame and at an affordable cost. Furthermore, extensive examination of the mt genomes of large cohorts of control and patient populations will contribute largely to refine further the spectrum of rare variants responsible for mtDNA disease and unusual but clinical irrelevant nucleotide diversity. NGS analysis of the human genomes is definitively here to stay, but the rate at which this technology will conquer the diagnostic mt genome bench will largely depend on the power of the analytical and bioinformatic procedures.
We thank patients and physicians for their contribution to this research project. Eur J Hum Genet. Published online Mar Author information Article notes Copyright and License information Disclaimer. This article has been cited by other articles in PMC. Supplementary Table 2. Supplementary Table 3. Supplementary Table 4. Abstract Next-generation sequencing NGS , an innovative sequencing technology that enables the successful analysis of numerous gene sequences in a massive parallel sequencing approach, has revolutionized the field of molecular biology.
Introduction Mitochondrial disorders are a very complex and heterogeneous group of human diseases, affecting multiple organs or tissues, and are caused by malfunction of the cellular energy production system hosted by the mitochondria. Preparation of artificial heteroplasmic samples to determine the detection limit of different bases at the same nucleotide position Combining PCR products, and not gDNA of Control samples 2 and 3 Supplementary Table 1 , is a prerequisite to this experiment, as the copy numbers of mtDNA molecules vary substantially among individuals.
Data analysis Reads were generated using the Ion Torrent platform pipeline software v. Open in a separate window. Figure 1. Sequencing of the pUC19 plasmid to define error rate Base calling errors associated with NGS data could result in false-positives as they, incorrectly, might be considered as heteroplasmic nucleotide variants. Determination of the detection limit of mtDNA heteroplasmy in patient samples To determine both the accuracy and precision of our settings for identification of heteroplasmy and for evaluation of the reliability of quantitative measurements, a DNA control sample was spiked in with another control DNA.
Figure 2. Variant detection using the Torrent Suite 3. Table 1 Overview of all pathogenic and heteroplasmic variants. PhePro Leukocytes 11 No Yes 2. LeuPro Leukocytes 18 No Yes 0. IleThr Leukocytes 25 Yes 1. IleThr Leukocytes 9 No No 3. Ile36Thr Leukocytes 8 No No 0. AspAsn Muscle 80 Yes 6.
Detection of three novel potential pathogenic variants In our data set, three novel alterations with a potential pathogenic character were detected with both MPS and conventional sequencing. Heteroplasmic variant calling A total of 25 heteroplasmic variants at 23 different positions m.
Detection of large single or multiple deletions in patients This study population included two skeletal muscle samples Supplementary Table 1 with single large-scale deletions. Figure 3. Discussion Mutations of mtDNA are frequent in humans and implicated in a heterogeneous group of mt disease pathology. Acknowledgments We thank patients and physicians for their contribution to this research project.
Notes The authors declare no conflict of interest. Supplementary Material Supplementary Table 1 Click here for additional data file. Supplementary Table 2 Click here for additional data file. Supplementary Table 3 Click here for additional data file. Supplementary Table 4 Click here for additional data file.
Human mitochondrial DNA: roles of inherited and somatic mutations. Nat Rev Genet. Mitochondrial genetics. Br Med Bull. Factors affecting the detection and quantification of mt point heteroplasmy using Sanger sequencing and SNaPshot minisequencing. Int J Legal Med. Sequencing technologies — the next generation. Comprehensive NGS analyses of the entire mitochondrial genome reveal new insights into the molecular diagnosis of mitochondrial DNA disorders. Genet Med. PLoS One. Complete mitochondrial genome amplification.
Nat Genet. A novel sporadic mutation in cytochrome c oxidase subunit II as a cause of rhabdomyolysis. Neuromuscul Disord. Performance comparison of benchtop high-throughput sequencing platforms. Nat Biotechnol. Detecting heteroplasmy from high-throughput sequencing of complete human mitochondrial DNA genomes. Am J Hum Genet. Heteroplasmic mitochondrial DNA mutations in normal and tumour cells. Dynamics of mitochondrial heteroplasmy in three families investigated via a repeatable re-sequencing study.
Genome Biol. Universal heteroplasmy of human mitochondrial DNA. Hum Mol Genet. PLoS Comput Biol. A mtDNA microdeletion in a newborn girl with transient lactic acidosis. J Inherit Metab Dis. Pediatr Res. Book Description: Chemistry is designed to meet the scope and sequence requirements of the two-semester general chemistry course.
The textbook provides an important opportunity for students to learn the core concepts of chemistry and understand how those concepts apply to their lives and the world around them. The book also includes a number of innovative features, including interactive exercises and real-world applications, designed to enhance student learning.
Appendix A: The Periodic Table. Appendix B: Essential Mathematics. Appendix C: Units and Conversion Factors. Appendix D: Fundamental Physical Constants. Appendix E: Water Properties. Appendix J: Solubility Products. Chemistry is designed to meet the scope and sequence requirements of the two-semester general chemistry course. Under this license, any user of this textbook or the textbook contents herein must provide proper attribution as follows:. For questions regarding this license, please contact partners openstaxcollege.
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If darts are closed to each other, then it is high precision. Similarly, when they are spread out, it is low precision. In the above figure, the four possible conditions are illustrated. In laboratories, the regular calibration of instrumentations is necessary to maintain the accuracy and precision of measurements.
Consider an experiment involving measuring the mass of copper sulphate whose actual weight is 2. Student A measures the mass of the salt thrice on weighing machine, he gets values 2. The same measurements are repeated by student B, student C, and student D on different weighing machines. All the readings are reported in the table below. From the above table, we can conclude that student A has performed the experiment correctly and precisely. For student B, it might be the weighing machine is not properly calibrated.
Student C is not following proper steps while taking the reading like not using vented balance safety enclosures. Finally, student D is the worst case of all the above. Accuracy vs Precision Darts throwing game When darts are landed on the dartboard away from the target point, we say it has low accuracy.
When darts are closed to each other as well as to the target point, it is high precision and high accuracy. This is the desired condition. Chemists always try to get results with great precision and accuracy. In 2 nd condition, all the darts are closed to each other but away from the target.
Such a condition has low accuracy and high precision. When talking about the accuracy of a measurement, Chemists like to know just how accurate the measurement is. For this reason, we calculate the percentage relative error as shown below:. The iron cube is weighed on 4 different electronic balances, and the percentage relative error is calculated for each measurement as shown below:.
When balance 1. When balance 2. If we know the tolerance of a true measurement, we can decide that the determined value is accurate if it lies within the tolerance levels of the true measurement, and, inaccurate if it lies outside the tolerance levels of the true value. For example, the cube of iron has a true value of 7.
An accurately determined value for the mass of iron would be between 7. An inaccurately determined value for the mass of iron would be less than 7. In order to determine how accurate a measurement is, we need to know the true value. Often we don't know the true value, so we can't determine how accurate our results are.
Play the game now! Because it is very rare for Chemists to know how accurate a measurement is, they make a number of measurements under the same conditions until they arrive at a set of measurements that are in good agreement with each other. The reproducibility of a measurement is known as precision. When all the measurements are very similar, we say the determined value is known precisely.
If we cannot get measurements that are very similar, we cannot say the value is known precisely, instead we say the measurements are imprecise. We can make generalisations about the precision of a set of measurements by calculating the range of the experimentally determined values:. A set of measurements can be described as precise if the range of values is very small, that is, range is close to 0 A set of measurements will be described as imprecise if the range of values is large, that is, range is not close to 0.
Example: The mass of the iron cube was measured 3 times in three separate trials using Balance 1 above. The results are shown below:. Using Balance 1. Balance 1. Example: The mass of the iron cube was measured 3 times in three separate trials using Balance 2 above. Using Balance 2. Balance 2. Take the test now! Example 1. The temperature of a laboratory is known to be a constant A student uses an alcohol thermometer to measure the temperature of the laboratory three times and records the temperatures in the table below:.
Deciding if the Student's results are accurate. To determine accuracy, we need to compare each of the student's results to the true value given as Given that we have a number of different measurements using the same thermometer by the student in the same laboratory, we could start by averaging the student's results:.
Next, we can use this average value to calculate the percentage relative error by allowing the average recorded temperature to be the determined value:. The student's temperature readings are NOT accurate, they are inaccurate. Deciding if the Student's results are precise.
Accuracy and precision in chemistry pdf torrent james chance and the contortions torrentAccuracy, precision, significant figures, mean and lara.torentinojum.space concepts of analytical chemistry.
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