Arichard:Notebook/2013/December: Difference between revisions
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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). | 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). | ||
===12/11/2013=== | |||
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[[File: 2013_12_11_bulk_precut1.jpg|400px]] | |||
[[File: 2013_12_11_bulk_precut2.jpg|400px]] | |||
[[File: 2013_12_11_bulk_cut1.jpg|400px]] | |||
[[File: 2013_12_11_bulk_cut2.jpg|400px]] |
Revision as of 00:46, 12 December 2013
December, 2012
12/02/2013
Meeting with Ben Siddoway and Gwen Kaeser.
Discussed possible quality control measures.
- Neuronal nuclei come from three brain banks:
- UC Irvine
- Maryland
- Wash U
12/03/2013
Shadowed Gwen for neuronal nuclei isolation
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).
12/11/2013
File:2013 12 11 bulk nxtpcr.jpg
File:2013 12 11 bulk precut1.jpg