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Figure 1. Comparison of cfDNA recovery using different commercially available kits.

A. Average cfDNA yield per 1 ml of plasma with 4 extraction kits, quantified using qPCR for GAPDH , ACTBL2 and HPRT and Qubit® 2.0 High Sensitivity dsDNA Assay. Error bar = ±SEM, n = 9 from 3 independent experiments each performed in triplicate. ND = Not detectable. B. Percent recovery of serial dilutions of λ/ Hind III DNA, quantified using assays targeting the 23 kb and 564 bp fragments. Error bar = ±SEM, n = 9 from 3 independent experiments performed in triplicate. CNA = Circulating Nucleic Acid kit.

https://doi.org/10.1371/journal.pone.0077963.g001

To determine the efficiency of cfDNA recovery, we assessed recovery of spiked-in λ/ Hind III DNA using the 3 kits that gave the highest yields of cfDNA (QIAamp ® DNA Blood Mini Kit, QIAamp ® CNA Kit and NucleoSpin ® Plasma XS Kit). The QIAamp ® DNA Blood Mini kit and the QIAamp ® CNA Kit showed similar but variable recovery of the 23kb fragment (range 20-70%). The QIAamp ® CNA Kit gave the highest yield of the smaller (564 bp) fragment, recovering between 90 - 100% at each dilution tested; whereas the NucleoSpin ® Plasma XS Kit showed a lower recovery of both amplicons ( Outlet Shop Offer Santoni Leather brogue loafers Outlet 100% Guaranteed Buy Cheap Latest Wide Range Of Online Discount Fashion Style F3GKmgsc
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To determine whether the two kits giving the best recovery of cfDNA (QIAamp ® DNA Blood Mini kit and the QIAamp ® CNA Kit) showed any variance in cfDNA profiles, paired cfDNA from 4 metastatic patients were sequenced using the Ion AmpliSeq™Cancer Hotspot Panel v2 on the Ion PGM™ Sequencer. A total of 412M bases were generated across the eight cfDNA samples, with an average of 52 ± 5 M bases per sample ( Sale Footlocker Finishline SAN FRANCISCO GTX Walking trainers dunkelbraun Cheap Sale Many Kinds Of Visit New Cheap Price Reliable For Sale eea9QTspWS
). This was equivalent to 503 ± 46K reads, > 96% of which mapped to the target region providing an average coverage of 2000x. High similarity was observed in the coverage profile for each cfDNA pair across the 207 amplicons (average size 154 bp, range 111bp -187 bp) ( Cheap Sale Professional Jack Rogers Kane Plaid Authentic For Sale Cheap Factory Outlet Visa Payment For Sale 30bzyR
). Of note, one sample (P4) showed a peak in the number of reads (>30,000 reads) at ERBB2 suggesting gene amplification. The total number of reads at each amplicon showed high correlation, (ρ > 0.95, p <0.001) for 3 of the 4 cfDNA pairs ( Footlocker Pictures Sale Online Buy Cheap Sast Marcelo Burlon logo detail espadrilles Black Ol3UW77q
). In the one sample (P3) with lower but significant correlation (ρ = 0.87) the QIAamp ® CNA method gave a more uniform coverage that the QIAamp ® DNA Blood Mini method.

Figure 2. Comparison of number of reads between two different extraction kits using the Ion AmpliSeq™Cancer Hotspot Panel v2.

Analysis of 4 plasma samples extracted using both the QIAamp ® DNA Blood Mini kit and the QIAamp ® CNA Kit. In each trace, the number of reads is shown on the Y-axis. Red lines indicate the QIAamp ® CNA Kit and the blue lines represent the QIAamp ® DNA Blood Mini kit. For in depth analysis see adidas Cloudfoam Super Racer Outlet Comfortable Cheap Sale Discount O8XTuv7QE
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Some 40% of companies now try to fight bias with mandatory hiring tests assessing the skills of candidates for frontline jobs. But managers don’t like being told that they can’t hire whomever they please, and our research suggests that they often use the tests selectively. Back in the 1950s, following the postwar migration of blacks northward, Swift Company, Chicago meatpackers, instituted tests for supervisor and quality-checking jobs. One study found managers telling blacks that they had failed the test and then promoting whites who hadn’t been tested. A black machine operator reported: “I had four years at Englewood High School. I took an exam for a checker’s job. The foreman told me I failed” and gave the job to a white man who “didn’t take the exam.”

This kind of thing still happens. When we interviewed the new HR director at a West Coast food company, he said he found that white managers were making only strangers—most of them minorities—take supervisor tests and hiring white friends without testing them. “If you are going to test one person for this particular job title,” he told us, “you need to test everybody.”

But even managers who test everyone applying for a position may ignore the results. Investment banks and consulting firms build tests into their job interviews, asking people to solve math and scenario-based problems on the spot. While studying this practice, Kellogg professor Lauren Rivera played a fly on the wall during hiring meetings at one firm. She found that the team paid little attention when white men blew the math test but close attention when women and blacks did. Because decision makers (deliberately or not) cherry-picked results, the testing amplified bias rather than quashed it.

Managers made only strangers—most of them minorities—take tests and hired white friends without testing them.

Companies that institute written job tests for managers—about 10% have them today—see decreases of 4% to 10% in the share of managerial jobs held by white women, African-American men and women, Hispanic men and women, and Asian-American women over the next five years. There are significant declines among white and Asian-American women—groups with high levels of education, which typically score well on standard managerial tests. So group differences in test-taking skills don’t explain the pattern.

More than 90% of midsize and large companies use annual performance ratings to ensure that managers make fair pay and promotion decisions. Identifying and rewarding the best workers isn’t the only goal—the ratings also provide a litigation shield. Companies sued for discrimination often claim that their performance rating systems prevent biased treatment.

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that raters tend to lowball women and minorities in performance reviews. And some managers give everyone high marks to avoid hassles with employees or to keep their options open when handing out promotions. However managers work around performance systems, the bottom line is that ratings don’t boost diversity. When companies introduce them, there’s no effect on minority managers over the next five years, and the share of white women in management drops by 4%, on average.

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