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===12/03/2013=== Shadowed Gwen for neuronal nuclei isolation [[http://genome-tech.ucsd.edu/LabNotes/index.php/Arichard:Protocols/neuronalnucleiisolation20131203| Protocol used]] We decided to use the same samples, but take samples along the way for bulk prep (Nextera). Female preferred so that presence of Y chromosome can be used as a marker for contamination by myself. Bulk sorted 3 types, all NeuN+ * 1864+, female Down's from ???. * 1568+, female AD from Maryland. * 61788+, female AD from Wash U. (New patient) Notes * Started with 0.095 g 1568, 0.14 g 61788, and 0.3 g 1864. * Filtered with 40 um BD Falcon filter. * Fixed 25 min, not 30. Time point samples (3) for bulk prep on each sample (3) (9 total): * 1 = After filter * 2 = Before FACS * 3 = After FACS All samples were stained with PI instead of DAPI due to concerns about the DNA damage upon excitation. This was unnecessary, however, as DAPI uses a 405 nm Ex wavelenth, which is not UV and nowhere near the absorption peak for thymine dimerization (254 nm).
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