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The positive transformants were screened on MD medium and analyzed by PCR.
(D) Atg31 protein was isolated by GST-tag purification from yeast grown in full or SD-N medium and analyzed by mass spectrometry.
Coverslips have been mounted in Dako mounting medium and analyzed with a Leica confocal microscope.
Embryos were washed in embryo medium and analyzed under a fluorescence microscope.
Coverslips were mounted on microscope glass slide with DAKO mounting medium and analyzed using immunofluorescent confocal microscopy or laser scanning cytometry (CompuCyte Corp), essentially as previously described [28].
Microfossil slides were prepared using permanent mounting medium and analyzed using a Nikon microscope with phase-contrast illumination at 1000× magnification [29], [30].
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We therefore investigated the stability of the genomic rearrangements by isolating six additional clones, called BW4005.C1 to BW4005.C6, after plating the initial glycerol stock of BW4005 onto LB medium, and analyzing them by RFLP-IS 3 and PFGE.
Slides were then mounted in antifade mounting medium (Ultracruz mounting medium, Santacruz) and analyzed with a Zeiss LSM 510 META confocal laser scanning microscope (Zeiss, Thornwood, NY) using LSM Image Examine software.
For analysis of proMMP-2 activation, conditioned medium was collected after 24 hours in serum-free medium, concentrated, and analyzed for gelatinase activity by zymography.
Finally, coverslips were mounted on glass slides with Vectashield mounting medium (Vector Laboratories) and analyzed by Confocal Microscopy (Leica DMIRE2 and TCS-SPE).
Sections and coverslides were mounted in anti-fade mountant medium (Sigma) and analyzed by Confocal laser microscope (LSM 510 version 2.01; Ziess, Thornwood, NY).
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