Daniel:Notebook/HiResChrPaint/2014-1-27: Difference between revisions

From ZhangLabWiki
Jump to navigation Jump to search
>Djacobse
(Created page with "=FISH Slide Prep= Back to Calendar I've got cells ready to split so I'm making some dishes for FISH. ==FISH Slide Preparation== ''Origina...")
 
>Djacobse
 
Line 1: Line 1:
=FISH Slide Prep=
=Alu Control FISH ([[Daniel:Notebook/HiResChrPaint/2014-1-24#FISH|Started 01/24/2014]])=


[[Daniel:Notebook/HiResChrPaint|Back to Calendar]]
[[Daniel:Notebook/HiResChrPaint|Back to Calendar]]


I've got cells ready to split so I'm making some dishes for FISH.
==Microscopy Results==
 
==FISH Slide Preparation==
 
''Original Protocol from Beliveau et al'': [[File:Slide_preparation_for_interphase_FISH.pdf]]
 
*Prepared 4 dishes PGP1F P9
 
1. Prepare a cell suspension of 500 000-2 000 000 cells/mL in growth media
2. Add 100 uL of cell suspension per dish roughly in the center
3. Allow cells to adhere for 3 hours at the 37 C
4. Rinse briefly in 1X PBS
5. Fix slides for 5-15 minutes in 4% (v/v) paraformaldehyde in 1X PBS
6. Rinse briefly in 1X PBS
7. Incubate for 5 minutes in 2X SSCT
8. Incubate for 5 minutes in 2X SSCT + 50% (v/v) formamide
9. Add fresh 2X SSCT + 50% (v/v) formamide for storage at 4C
 
===Pictures===
 
I took snapshots of all four dishes after fixing, just to be sure the cells were still there (since I've had trouble finding cells previously).
 


{| class="wikitable"
{| class="wikitable"

Latest revision as of 20:36, 27 January 2014

Alu Control FISH (Started 01/24/2014)[edit]

Back to Calendar

Microscopy Results[edit]

File:2014 01 27 Cot1 Positive Control Nuc1.jpg|File:2014 01 27 Cot1 Positive Control Nuc2.jpg|
Two nucleii stained with Draq5 (red) and Alexa 488 (Cot1 DNA, green)
File:2014 01 27 V6S3 488.jpg|
Nucleus stained with Draq5 (red) and Alexa 488 (V6S3, green)