PALM Protocols – RNA handling
Dos and Don’ts, slide preparation, archived samples, staining procedures, storage, laser microdissection procedure, collection devices and procedures. Downstream applications, RNA from FFPE and frozen sections, QIAGEN RNeasy FFPE kit, quality control, lab tips.
(28 pages)
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PALM Protocols – DNA handling
Preparation and treatment of slides, mounting samples onto slides, staining procedures, laser microdissection procedure, collection devices and procedures, cap check. Downstream applications, DNA from FFPE and frozen sections. QIAamp DNA procedure. PCR set up.
(26 pages)
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Chromosome Preparation
Chromosome preparation, staining, procedure of laser microdissection, collection procedures, collection onto AmpliGrid slide, consumables and components. Collection devices, lenses.
(20 pages)
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Cell Culture, non-contact Laser Capture Microdissection (LCM) and Recultivation
Cultivation of cells, coating of Dishes and Slides, improvement of visualization, procedure of laser microdissection, recultivation, time lapse, incubators. Optical Tweezers.
(14 pages)
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Immunofluorescence on frozen sections
Tips before starting, microtome cutting, antibody staining.
(5 pages)
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FISH Hybridization
Chromosome preparation. Preparation of chromosome paints. Hybridization of painting probes. Preparation of centromeric probes.
(15 pages)
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AutoLPC from glass slides results in good quality RNA
For RNA experiments on microdissected samples we usually recommend the use of MembraneSlides. Sometimes only old archived slides are available. We show that even this kind of slides is useful for RNA preparation. After removing the coverslip the unique and proprietary software function of “AutoLPC” allows quick access to the desired tissue area. And we could prove that this method does not influence the RNA quality.
(6 pages)
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RNA extraction from FFPE sections
Short protocol. Tips and recommendations for using FFPE sections as source for RNA extraction.
(4 pages)
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RNA extraction from frozen sections
Short protocol. Especially helpful for RNase rich tissue. Tips for microtome cutting and special staining (cresylviolet).
(4 pages)
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