Daniel:Notebook/HiResChrPaint/2014-2-13: Difference between revisions

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==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" | &nbsp;
| 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" | &nbsp;
| 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]

Back to Calendar

Cot1 Labeling (Started 02/12/2014)[edit]

  1. Remove samples from -80C and centrifuge for 20 minutes at 12000 RPM at 4 C
  2. Remove supernatant and replace with 750uL 75% EtOH
  3. Centrifuge for 10 minutes at 12000 RPM at 4 C
  4. Remove supernatant and leave in hood to dry
  5. Resuspend each sample in 20 uL nfH2O
    1. Measure concentration in nanodrop
    2. Run on TBE gel to ensure correct size distribution
  6. 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

File:2013-02-13-HiLoMW.jpg

Pooled Nanodrop Results

  High MW Low MW
ng/uL 12.2 20.3
ug in ~60 uL 0.7 1.2