Daniel:Notebook/HiResChrPaint/2014-2-13: Difference between revisions
Jump to navigation
Jump to search
>Djacobse (Created page with "=Multi-Color FISH (Started 02/08/2014)= Back to Calendar ==Cot1 Labeling ([[Daniel:Notebook/HiRe...") |
>Djacobse No edit summary |
||
(6 intermediate revisions by the same user not shown) | |||
Line 4: | Line 4: | ||
==Cot1 Labeling ([[Daniel:Notebook/HiResChrPaint/2014-2-12|Started 02/12/2014]])== | ==Cot1 Labeling ([[Daniel:Notebook/HiResChrPaint/2014-2-12|Started 02/12/2014]])== | ||
#Remove samples from -80C and centrifuge for 20 minutes at 12000 RPM at 4 C | |||
#Remove supernatant and replace with 750uL 75% EtOH | |||
#Centrifuge for 10 minutes at 12000 RPM at 4 C | |||
#Remove supernatant and leave in hood to dry | |||
#Resuspend each sample in 20 uL nfH2O | |||
##Measure concentration in nanodrop | |||
##Run on [[Daniel:Protocols/TBEgel|TBE gel]] to ensure correct size distribution | |||
#Pool all high MW samples and all low MW samples | |||
===Results=== | |||
'''Nanodrop Results''' | |||
{| 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="15" | High MW | |||
| width="65" | A | |||
| width="65" | B | |||
| width="65" | C | |||
| width="65" | D | |||
| width="65" | Low MW | |||
| width="65" | A | |||
| width="65" | B | |||
| width="65" | C | |||
| width="65" | D | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | ng/uL | |||
| align="center" | 10.3 | |||
| align="center" | 44.8 | |||
| align="center" | 13.6 | |||
| align="center" | 10.1 | |||
| align="center" | | |||
| align="center" | 17.7 | |||
| align="center" | 20.4 | |||
| align="center" | 22.5 | |||
| align="center" | 70.9 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | ug in 20 uL | |||
| align="center" | 0.2 | |||
| align="center" | 0.9 | |||
| align="center" | 0.3 | |||
| align="center" | 0.2 | |||
| align="center" | | |||
| align="center" | 0.4 | |||
| align="center" | 0.4 | |||
| align="center" | 0.5 | |||
| align="center" | 1.4 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | Total | |||
| align="center" | | |||
| align="center" | | |||
| align="center" | | |||
|style="font-weight:bold" align="center" | 1.6 | |||
| align="center" | | |||
| align="center" | | |||
| align="center" | | |||
| align="center" | | |||
|style="font-weight:bold" align="center" | 2.6 | |||
|} | |||
'''Gel Image''' | |||
{{GelLanes|Lane3=Low Mass Ladder|Lane4=High MW|Lane5=High MW|Lane6=High MW|Lane7=High MW|Lane8=Low MW|Lane9=Low MW|Lane10=Low MW|Lane12=Low MW}} | |||
[[Image:2013-02-13-HiLoMW.jpg|500px]] | |||
'''Pooled Nanodrop Results''' | |||
{| 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" | |||
| align="center" width="65" height="15" | | |||
| width="65" | High MW | |||
| width="65" | Low MW | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | ng/uL | |||
| align="center" | 12.2 | |||
| align="center" | 20.3 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | ug in ~60 uL | |||
| align="center" | 0.7 | |||
| align="center" | 1.2 | |||
|} | |||
[[Category:HRCP]] [[Category:20140208]] |
Latest revision as of 18:21, 2 July 2014
Multi-Color FISH (Started 02/08/2014)[edit]
Cot1 Labeling (Started 02/12/2014)[edit]
- Remove samples from -80C and centrifuge for 20 minutes at 12000 RPM at 4 C
- Remove supernatant and replace with 750uL 75% EtOH
- Centrifuge for 10 minutes at 12000 RPM at 4 C
- Remove supernatant and leave in hood to dry
- Resuspend each sample in 20 uL nfH2O
- Measure concentration in nanodrop
- Run on TBE gel to ensure correct size distribution
- Pool all high MW samples and all low MW samples
Results[edit]
Nanodrop Results
High MW | A | B | C | D | Low MW | A | B | C | D |
ng/uL | 10.3 | 44.8 | 13.6 | 10.1 | 17.7 | 20.4 | 22.5 | 70.9 | |
ug in 20 uL | 0.2 | 0.9 | 0.3 | 0.2 | 0.4 | 0.4 | 0.5 | 1.4 | |
Total | 1.6 | 2.6 |
Gel Image
Lane | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 |
Sample | Low Mass Ladder | High MW | High MW | High MW | High MW | Low MW | Low MW | Low MW | Low MW |
Pooled Nanodrop Results
High MW | Low MW | |
ng/uL | 12.2 | 20.3 |
ug in ~60 uL | 0.7 | 1.2 |