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===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. Denature DNA by incubating at 95 C for 5 minutes, followed by snap cooling on ice 4. Thaw out dyes (they are in big -20C) and resuspend in 2.2 uL DMSO Use dyes immediately, they do not store 5. Add 1 uL of appropriate dye to oligo set (see below) Chose V4S2 and V6S2 because of high dna content so I only had to use one dye (Alexa 546) {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> |- style="font-size:12pt" align="center" | width="65" height="16" | Probe | width="65" | Alexa Dye |- style="font-size:12pt" align="center" | height="16" | V4S1 | align="right" | 488 |- style="font-size:12pt" align="center" | height="16" | V4S2 | align="right" | 546 |- style="font-size:12pt" align="center" | height="16" | V4S3 | align="right" | 594 |- style="font-size:12pt" align="center" | height="16" | V4S4 | align="right" | 647 |- style="font-size:12pt" align="center" | height="16" | V6S1 | align="right" | 488 |- style="font-size:12pt" align="center" | height="16" | V6S2 | align="right" | 546 |- style="font-size:12pt" align="center" | height="16" | V6S3 | align="right" | 594 |- style="font-size:12pt" align="center" | height="16" | V6S4 | align="right" | 647 |} 6. Incubate reaction in the dark for 1 hour 7. Column purify using ssDNA kit from Zymo
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