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10.1373/clinchem.2015.249623 http://scihub22266oqcxt.onion/10.1373/clinchem.2015.249623 C4878677!4878677!26847218     free     free     free Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Clin+Chem 2016 ; 62 (4): 647-54 Nephropedia Template TP {{tp|p=26847218|t=2016. Systematic Evaluation of Sanger Validation of NextGen Sequencing Variants |pdf=|usr=}}{{26847218}} gab.com Text Systematic Evaluation of Sanger Validation of NextGen Sequencing Variants JO: Clin Chem http://www.kidney.de/pget.php?&tw=1&pmid=26847218 #FOAvip BACKGROUND: Next-generation sequencing (NGS) data are used for both clinical care and clinical research. DNA sequence variants identified using NGS are often returned to patients/participants as part of clinical or research protocols. The current standard of care is to validate NGS variants using Sanger sequencing, which is costly and time-consuming. METHODS: We performed a large-scale, systematic evaluation of Sanger-based validation of NGS variants using data from the ClinSeq® project. We first used NGS data from 19 genes in five participants, comparing them to high-throughput Sanger sequencing results on the same samples, and found no discrepancies among 234 NGS variants. We then compared NGS variants in five genes from 684 participants against data from Sanger sequencing. RESULTS: Of over 5,800 NGS-derived variants, 19 were not validated by Sanger data. Using newly-designed sequencing primers, Sanger sequencing confirmed 17 of the NGS variants, and the remaining two variants had low quality scores from exome sequencing. Overall, we measured a validation rate of 99.965% for NGS variants using Sanger sequencing, which was higher than many existing medical tests that do not necessitate orthogonal validation. CONCLUSIONS: A single round of Sanger sequencing is more likely to incorrectly refute a true positive variant from NGS than to correctly identify a false positive variant from NGS. Validation of NGS-derived variants using Sanger sequencing has limited utility, and best practice standards should not include routine orthogonal Sanger validation of NGS variants. Twit Text FOAVip Systematic Evaluation of Sanger Validation of NextGen Sequencing Variants ????Clin Chem http://www.kidney.de/pget.php?&tw=1&pmid=26847218 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=26847218 #FOAvip English Wikipedia <ref>{{Cite pmid|26847218}}</ref> Systematic Evaluation of Sanger Validation of NextGen Sequencing Variants #MMPMID26847218 Beck TF; Mullikin JC; Biesecker LG Clin Chem 2016[Apr]; 62 (4): 647-54 PMID26847218show ga BACKGROUND: Next-generation sequencing (NGS) data are used for both clinical care and clinical research. DNA sequence variants identified using NGS are often returned to patients/participants as part of clinical or research protocols. The current standard of care is to validate NGS variants using Sanger sequencing, which is costly and time-consuming. METHODS: We performed a large-scale, systematic evaluation of Sanger-based validation of NGS variants using data from the ClinSeq® project. We first used NGS data from 19 genes in five participants, comparing them to high-throughput Sanger sequencing results on the same samples, and found no discrepancies among 234 NGS variants. We then compared NGS variants in five genes from 684 participants against data from Sanger sequencing. RESULTS: Of over 5,800 NGS-derived variants, 19 were not validated by Sanger data. Using newly-designed sequencing primers, Sanger sequencing confirmed 17 of the NGS variants, and the remaining two variants had low quality scores from exome sequencing. Overall, we measured a validation rate of 99.965% for NGS variants using Sanger sequencing, which was higher than many existing medical tests that do not necessitate orthogonal validation. CONCLUSIONS: A single round of Sanger sequencing is more likely to incorrectly refute a true positive variant from NGS than to correctly identify a false positive variant from NGS. Validation of NGS-derived variants using Sanger sequencing has limited utility, and best practice standards should not include routine orthogonal Sanger validation of NGS variants. äSystematic Evaluation of Sanger Validation of NextGen Sequencing Varia(epmc).pdf DeepDyve Pubget Overpricing