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 - 06 Jun 2012 - 19:43:00 - Main.AlvaroRodriguez
 
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