Daniel:Notebook/HiResChrPaint/2014-2-23: Difference between revisions
Jump to navigation
Jump to search
>Djacobse (Created page with "=Multi-Color FISH (Started 02/08/2014)= Back to Calendar ==Hybridization== '''Sample Matrix''' ...") |
>Djacobse |
||
Line 7: | Line 7: | ||
'''Sample Matrix''' | '''Sample Matrix''' | ||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="font-size:12pt;font-weight:bold" align="center" | |||
| width="65" height="30" | Sample | |||
| width="91" | Sample A | |||
| width="65" | uL sample A (pmol) | |||
| width="65" | Sample B | |||
| width="65" | uL sample B (pmol) | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="30" | V6S3 Only | |||
| [[Daniel:Notebook/HiResChrPaint/2014-2-21#Labeling Results|V6S3-546]] | |||
| 5 (10) | |||
| Unlabeled Cot1 | |||
| 1 (1 ug) | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | V6S3+Cot1 | |||
| [[Daniel:Notebook/HiResChrPaint/2014-2-21#Labeling Results|V6S3-546]] | |||
| 5 (10) | |||
| [[Daniel:Notebook/HiResChrPaint/2014-1-24#Labeling Results|Cot1-488]] | |||
| 6 (10) | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="30" | Multi-Color Cot1 | |||
| [[Daniel:Notebook/HiResChrPaint/2014-2-21#Labeling Results|Cot1-High-546]] | |||
| 20 (4) | |||
| [[Daniel:Notebook/HiResChrPaint/2014-2-21#Labeling Results|Cot1-Low-488]] | |||
| 5 (5) | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="45" | BSPP Negative Control | |||
| [[Daniel:Notebook/HiResChrPaint/2014-2-21#Labeling Results|BSPP1]] | |||
| 5 (10) | |||
| Unlabeled Cot1 | |||
| 1 (1 ug) | |||
|} | |||
''Day 1'' | ''Day 1'' |
Revision as of 00:32, 23 February 2014
Multi-Color FISH (Started 02/08/2014)
Hybridization
Sample Matrix
Sample | Sample A | uL sample A (pmol) | Sample B | uL sample B (pmol) |
V6S3 Only | V6S3-546 | 5 (10) | Unlabeled Cot1 | 1 (1 ug) |
V6S3+Cot1 | V6S3-546 | 5 (10) | Cot1-488 | 6 (10) |
Multi-Color Cot1 | Cot1-High-546 | 20 (4) | Cot1-Low-488 | 5 (5) |
BSPP Negative Control | BSPP1 | 5 (10) | Unlabeled Cot1 | 1 (1 ug) |
Day 1
- Allow stored samples to warm to room temperature
- Incubate in 2X SSCT +50% (v/v) formamide for 2.5 minutes at 92 C in pre-warmed media
- Incubate in 2X SSCT + 50% formamide at 60C for 20 minutes
- Remove samples and allow to cool to room temperature
- Add 25 uL hybridization cocktail to center
- Add 22x22 #1.5 coverslip; seal with rubber cement
- Allow the rubber cement to air-dry for 5 minutes at room temperature
- Denature for 2.5 minutes at 92C; keep humid
- Transfer slides to a humidified chamber and hybridize overnight at 37C or 42C