Daniel:Notebook/HiResChrPaint/2013-4-15: Difference between revisions

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  5. Incubate reaction in the dark for 1 hour
  6. Column purify using ssDNA kit from Zymo
[[Daniel:Notebook/HiResChrPaint|Back to Calendar]]
[[Daniel:Notebook/HiResChrPaint|Back to Calendar]]

Revision as of 18:19, 15 April 2013

Dye Coupling Test and Gel Quantification

 Purpose:  Test dye coupling reaction, specifically the purification step, to ensure proper dye-coupled DNA is being captured.
 Two samples chosen from Gel Quantification (4/10/13). 
 Oligos coupled to appropriate dye.

Dye Coupling Reaction

 1. Thaw out oligos, sodium bicarbonate solution, DMSO
    Use thermo device on Noi's desk to heat DMSO, sodium bicarb to 42 C
 2. Add 2.5 uL DNA, 1.5 uL sodium bicarbonate to 0.2 mL eppendorf
 3. Thaw out dyes (they are in big -20C) and resuspend in 2.2 uL DMSO
    Use dyes immediately, they do not store
 4. Add 1 uL of appropriate dye to oligo set (see below)
Probe Alexa Dye
V4S1 488
V4S2 546
V4S3 594
V4S4 647
V6S1 488
V6S2 546
V6S3 594
V6S4 647
 5. Incubate reaction in the dark for 1 hour
 6. Column purify using ssDNA kit from Zymo

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