Daniel:Notebook/HiResChrPaint/2014-1-14: Difference between revisions
Jump to navigation
Jump to search
>Djacobse No edit summary |
>Djacobse |
||
Line 25: | Line 25: | ||
8. [[Daniel:Protocols/CentriSep|Centri-Sep column purification]] | 8. [[Daniel:Protocols/CentriSep|Centri-Sep column purification]] | ||
9. Measure concentration in Nanodrop | 9. Measure concentration in Nanodrop | ||
{| 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="65" | ng/uL dsDNA | |||
| width="65" | pmol/uL dye | |||
| width="71" | pmol/uL ds probe | |||
| width="71" | dye/100bp | |||
| width="65" | ng/uL ssDNA | |||
| width="65" | pmol/uL dye | |||
| width="71" | pmol/uL ss probe | |||
| width="71" | dye/100bp | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="30" | Alu2D-Pre Column | |||
| align="center" | 170 | |||
| align="center" | 115 | |||
| align="center" | | |||
| align="center" | | |||
| align="center" | 113.2 | |||
| align="center" | 116.6 | |||
| align="center" | | |||
| align="center" | | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="30" | Blank-Pre Column | |||
| align="center" | 157.4 | |||
| align="center" | 111 | |||
| align="center" | | |||
| align="center" | | |||
| align="center" | 103.3 | |||
| align="center" | 110.6 | |||
| align="center" | | |||
| align="center" | | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="30" | Cot1-Pre Column | |||
| align="center" | 150.9 | |||
| align="center" | 105 | |||
| align="center" | | |||
| align="center" | | |||
| align="center" | 102.1 | |||
| align="center" | 106.4 | |||
| align="center" | | |||
| align="center" | | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="30" | V6S3-Pre Column | |||
| align="center" | 160.3 | |||
| align="center" | 109.2 | |||
| align="center" | | |||
| align="center" | | |||
| align="center" | 109.6 | |||
| align="center" | 112.9 | |||
| align="center" | | |||
| align="center" | | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="30" | Alu2D-Post Column | |||
| align="center" | 24.4 | |||
| align="center" | 0.5 | |||
| align="center" | 0.4 | |||
| align="center" | 1.4 | |||
| align="center" | 12.6 | |||
| align="center" | 0.5 | |||
| align="center" | 0.4 | |||
| align="center" | 1.3 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="30" | Blank-Post Column | |||
| align="center" | 1 | |||
| align="center" | 0 | |||
| align="center" | 0.0 | |||
| align="center" | 0.0 | |||
| align="center" | 0.2 | |||
| align="center" | 0 | |||
| align="center" | 0.0 | |||
| align="center" | 0.0 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="30" | Cot1-Post Column | |||
| align="center" | 25.9 | |||
| align="center" | 1.7 | |||
| align="center" | 0.2 | |||
| align="center" | 13.3 | |||
| align="center" | 15.6 | |||
| align="center" | 1.3 | |||
| align="center" | 0.3 | |||
| align="center" | 8.4 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="30" | V6S3-Post Column | |||
| align="center" | 41.3 | |||
| align="center" | 0.6 | |||
| align="center" | 0.8 | |||
| align="center" | 0.6 | |||
| align="center" | 24.4 | |||
| align="center" | 0.3 | |||
| align="center" | 0.9 | |||
| align="center" | 0.3 | |||
|} |
Revision as of 19:12, 15 January 2014
Alu Control (Started 1/08/2014)
Dye Coupling Test (with Positive Control)
Repeating the dye coupling test one last time using a positive control as well.
- Following ARES Protocol
Samples
0. Vaccuum centrifuge DNA; resuspend in 2.5 uL nfH2O 1. Warm DMSO and bicarbonate labeling buffer and nuclease free water to room temperature 2. Add 1.5 uL of sodium bicarbonate buffer 3. Denature samples for 5 minutes at 95C, then snap cool using ice box 4. Dissolve 1 vial of reactive dye in 2 uL solvent; vortex ≥10 seconds *dye cannot be saved for later use. Use immediately! 5. Add 1 uL dye to sample 6. Incubate in the dark for 1 hour 7. Add 20 uL nfH20 to sample; Measure concentration in Nanodrop 8. Centri-Sep column purification 9. Measure concentration in Nanodrop
Sample | ng/uL dsDNA | pmol/uL dye | pmol/uL ds probe | dye/100bp | ng/uL ssDNA | pmol/uL dye | pmol/uL ss probe | dye/100bp |
Alu2D-Pre Column | 170 | 115 | 113.2 | 116.6 | ||||
Blank-Pre Column | 157.4 | 111 | 103.3 | 110.6 | ||||
Cot1-Pre Column | 150.9 | 105 | 102.1 | 106.4 | ||||
V6S3-Pre Column | 160.3 | 109.2 | 109.6 | 112.9 | ||||
Alu2D-Post Column | 24.4 | 0.5 | 0.4 | 1.4 | 12.6 | 0.5 | 0.4 | 1.3 |
Blank-Post Column | 1 | 0 | 0.0 | 0.0 | 0.2 | 0 | 0.0 | 0.0 |
Cot1-Post Column | 25.9 | 1.7 | 0.2 | 13.3 | 15.6 | 1.3 | 0.3 | 8.4 |
V6S3-Post Column | 41.3 | 0.6 | 0.8 | 0.6 | 24.4 | 0.3 | 0.9 | 0.3 |