C.B.S. Scientific ECU-040 - Manual (Page 11)

C.B.S. Scientific Electro-Eluter Concentrator 11
3.4 Procedure for Elution of DNA
1. Please read through directions for elution of proteins. Follow instructions 3-5 of Section 3.1
using the buffer suggestions below.
a. Buffer: 5mM Tris-HoAc pH 7.5, 1mM EDTA, Current: 100-150 Volts @ 10mA,
Duration: Minimum of 5 hours for large DNA (8-10 kbp); 1.5 hours for 300 bp.
b. Comparisons of recoveries after 5 hours to 18 hours are similar. Therefore, it does
not hurt to run overnight when running large DNA fragments.
2. All electroelution preps are extensively Phenol extracted after run (2x, 1 hour) followed by
chloroform ether extraction. This is to remove any contaminants including dissolved agarose.
Note: Most people use a Brom-phenol-Blue Sucrose marker in with the piece of agarose to
make sure current flow is unobstructed by bubbles. After loading sample into chamber, drop
a little of this marker into the bottom of the loading chamber. Watch for movement of dye
after power is turned on.
3. DNA recovered from our ECU units has been tested for a number of viable characteristics
including kinase treatment, re-restriction, and ability to transform E. Coli and has been found
to be more active than DNA recovered by conventional techniques. Also, the percent
recoveries of DNA and the percent of specific E. Coli transformation by that DNA are much
better than traditional methods, despite extensive Phenol extraction.