Daniel:Notebook/HiResChrPaint/2013-10-25: Difference between revisions

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===Labeling Results===
===Labeling Results===
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext>
|- style="font-size:12pt" align="center"
| align="center" width="65" height="30" | &nbsp;
|style="font-weight:bold" width="65" | ng/uL DNA
|style="font-weight:bold" width="65" | pmol/uL dye
|style="font-weight:bold" width="65" | ug DNA in 20 uL
|style="font-weight:bold" width="65" | pmol/uL DNA
|style="font-weight:bold" width="71" | dye/probe
|style="font-weight:bold" width="71" | dye/100 bp
|style="font-weight:bold" width="71" | Base:Dye Ratio (:1)
|- style="font-size:12pt" align="center"
|style="font-weight:bold" height="15" | BSPP 1-546
| align="center" | 52
| align="center" | 1.7
| align="center" | 1.0
| align="center" | 1.9
| align="center" | 0.9
| align="center" | 1.1
| align="center" | 93
|- style="font-size:12pt" align="center"
|style="font-weight:bold" height="15" | BSPP 2-594
| align="center" | 68.8
| align="center" | 2.6
| align="center" | 1.4
| align="center" | 2.6
| align="center" | 1.0
| align="center" | 1.2
| align="center" | 80
|}

Revision as of 22:54, 25 October 2013

BSPP Control (Started 10/22/2013)

Back to Calendar

Dye Coupling

Just like with 9/03/2013 I'll be coupling BSPP-1 to Alexa 546 and BSPP-2 to Alexa 594

0. Dry samples in a vaccuum centrifuge

For each

1. Warm DMSO and bicarbonate labeling buffer and nuclease free water to room temperature
2. Resuspend DNA in 5 uL nfH20
3. Add 5 uL DNA and 3 uL of sodium bicarbonate buffer in PCR tube
4. Denature samples for 5 minutes at 95C, then snap cool using ice box
5. Dissolve 1 vial of reactive dye in 2 uL solvent; vortex ≥10 seconds
   *dye cannot be saved for later use.  Use immediately!
6. Add 2 uL dye to sample
7. Incubate in the dark for 1 hour
8. Add 10 uL nfH20 to sample
9. Purify using Centri Sep column
10. Measure concentrations using Microarray setting in nanodrop

Labeling Results

  ng/uL DNA pmol/uL dye ug DNA in 20 uL pmol/uL DNA dye/probe dye/100 bp Base:Dye Ratio (:1)
BSPP 1-546 52 1.7 1.0 1.9 0.9 1.1 93
BSPP 2-594 68.8 2.6 1.4 2.6 1.0 1.2 80