Arichard:Notebook/2013/December: Difference between revisions

From ZhangLabWiki
Jump to navigation Jump to search
>Andrew
>Andrew
No edit summary
Line 36: Line 36:


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===
[[File: 2013_12_11_bulk_nxtpcr.jpg|400px]]
[[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

[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).

12/11/2013

File:2013 12 11 bulk nxtpcr.jpg

File:2013 12 11 bulk precut1.jpg

File:2013 12 11 bulk precut2.jpg

File:2013 12 11 bulk cut1.jpg

File:2013 12 11 bulk cut2.jpg