Daniel:Notebook/HiResChrPaint/2013-7-2
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S2 Probe Prep (Started 5/31/2013)[edit]
Dye Coupling[edit]
Trying dye coupling today, followed by purification using Princeton Separations column. Princeton Sep column uses size exclusion instead of an affinity style column like Qiaquick or ssDNA (Zymo). Only using V4 sample as a trial run.
V4S2: ~1.2 ug
1. Warm reagents 2. Resuspend DNA in 5 uL nfH20 3. Add 3 uL sodium bicarbonate 4. Denature 5 minutes at 95C 5. Resuspend Alexa 488 dye in 2 uL DMSO 6. Add dye to sample 7. Incubate 1 hour in the dark 8. Size exclusion column
Results[edit]
ng/uL | ug in 15 uL | pmol/uL DNA | pmol/uL dye | fluorophores/ probe |
190.1 | 2.9 | 7.1 | 6.5 | 0.9 |
Seems like this is okay results. Some of Noi's results had this result, so it could be useful for FISH.