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Most of the available MRSA screening tests are based either on the identification of colony material grown on solid agar media or on direct molecular approaches.
In each protocol, colony material was scraped and the bacteria resuspended to allow inoculation of fresh plates in a transfer regime and/or to enumerate WS numbers.
A 1.0 µl inoculating loop was used to transfer B. anthracis colony material into 200 µl of Brain-Heart Infusion broth (Hardy Diagnostics, Santa Maria, CA) within the wells of a sterile, untreated polycarbonate 96-well culture plate (Costar Corning Inc., Acton, MA).
Gram stained colony material was examined and cell morphology recorded.
One loopful of colony material was used for DNA preparation.
One loopful of colony material was collected for DNA preparation.
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One micro litre of colony materials from the opaque zone of the colonies was streaked on XLD + N and BG, and incubated at 37 ± 1°C for 24 ± 3 h.
In 38% (±6%) of the wild-type cells, all ERCs that had accumulated in the mother cell remained in the mother cell, as demonstrated by the fact that their daughters lacked the ADE2 marker and formed a red colony (see material and methods, (Sinclair and Guarente, 1997)).
At the deep central and peripheral depressions areas of the ring and rough colonies extracellular material was seen in different arrangements.
Deep-layer CTIP2+ neurons migrated downward from the colonies towards the bottom of the collagen-filled space, with only a few CTIP2+ neurons present on the surface of the colonies (supplementary material Fig. S2).
As part of the assessment of pluripotency, we examined endogenous gene and protein expression of various markers associated with pluripotency including POU5F1, NANOG, SALL4 and DNMT3B in both OSKM and OSKMV colonies (Supplementary Material, Fig. S3C).
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