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In primary screening that considered title and abstract (if there was an abstract), we identified 208 potentially relevant articles and subsequently requested the full texts, of which 196 were accessible.
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Confirmatory analyses are required for the other 228 equally promising candidates identified in the primary screen that we have not yet tested in primary macrophages (marked by asterisks in Tables S1, S2, S3, S4).
Targets from the primary screen that produced glycogen staining phenotypes deviant from controls in terms of either density or localization were cloned into a new sublibrary and this library was rescreened three additional times in a blinded manner.
In the primary screen that involved the use of radiolabeled protein pools, both candidates were stable in the presence of XB and CycBΔ90 (mitotic extract containing activated APCCdc20), but they degraded in Xenopus egg extract supplemented with Cdh1, suggesting that they are substrates of APCCdh1 and not APCCdc20.
As in classical genetics, these large-scale HCS experiments are best considered primary screens that help in the discovery process.
Since a variety of factors, including promiscuous aggregators, non-selective covalent modifiers and compounds that sequester substrate molecules, can produce primary screening hits that are not relevant [44], we developed and implemented a panel of secondary assays to run against a subset of initial hits or the entire LOPAC collection.
Therefore, a simple high-throughput primary screening strategy that can culture and evaluate a large number of isolates simultaneously with a fair degree of accuracy and reproducibility becomes imperative [10].
In this evaluation of six HPV tests from residual liquid-based screening cytology specimens, all tests except for NorChip showed high sensitivity for high-grade lesions that were positive by cytology, suggesting that they are suitable for primary screening and that dual co-testing with cytology as well is unnecessary.
Images from the primary screen indicated that KD of GRINA in HeLa cells expressing CD8-CIMPR and GFP-GOLPH3 leads to a reduction in anti-CD8 antibody that reaches the TGN after a 30 min chase period.
The results from the primary screen indicated that siRNAs targeting a number of genes in the BER pathway decrease HIV infection.
Secondary screening of selected compounds proceeded similar to the primary screen except that the 10- and 100-fold lower concentrations of the effective compounds identified in the initial screens were tested both at 15°C and 6°C.
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Justyna Jupowicz-Kozak
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