---+++ *ELUTION of OLIGOS FROM PAGE GEL - Crush Soak* Eluting DNA/RNA from PAGE or denaturing PAGE ---++Materials Crush Soak Buffer (CSB) - 200mM NaCl , 10 mM tris-HCl (pH 7.5), 1mM EDTA (pH 8) using MilliQ water/NanoPure water. ---++Procedure (Work in Progress) Elution * 1 Excise Band (try doing it with the least polyacrylamide from gel need to get whole band) with a clean razor. * 2 Transfer the excised band into an 1.7 microliter eppi tube and crush into big chunks with a sterile pipet tip. * 3 Add 2 volumes of CSB for about every 1 volume of polyacrylamide. * 4 Incubate overnight at 37 C and by shaking it. (Speeds the process) * After the incubation, centrifuge the sample at 15000 rpm for 2 minutes at 4 C. * Trasfer supernatant(has the DNA/RNA) _carefully_ without aspirating any of the polyacrylamide to a new eppi tube. * *2X* Add 2 volumes of CBS, repeat centrifugation step, and transfer the supernatant. * Ethanol Precipitate the DNA from the supernatant.
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Topic revision: r3 - 2012-06-06 - AlvaroRodriguez
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