Efficient, small scale electroelution of high molecular weight DNA from agarose gels by a miniature vertical electrophoresis cell.
A miniature, vertical electrophoretic system is described that allows quick elution and efficient harvesting of double-stranded, high molecular weight DNA fragments from agarose gels. A gel slice containing the band of interest is excised from the agarose gel after submarine electrophoresis and placed into an electrophoresis cell, assembled by inserting a truncated micropipette tip into a 2.0 mL polypropylene tube. The tip supports the agarose slice and is connected to dialysis tubing. Platinum wire electrodes are placed over the tip and at the bottom of the test tube. DNA molecules are quickly eluted into the dialysis bag (60 s to 10 min for bands ranging from 2 to 23 kb) and then easily recovered. The method is highly efficient and allows considerable time saving. Of special interest is its applicability to small starting amounts of DNA (less than 0.5 micrograms per band). Purity of the DNA is comparable, if higher, to that obtainable by conventional electroelution on dialysis bags and DEAE-cellulose membrane electrophoresis. The system is simple enough to be used as a routine method for eluting multiple bands in short times.
Pascali
V L
VL
Immunohematology Laboratory, Istituto di Medicina Legale, Università Cattolica del S. Cuore, Roma, Italy.
Pescarmona
M
M
Dobosz
M
M
d'Aloja
E
E
D010582:Bacteriophage lambda; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification*; D004279:DNA, Viral / Q000302:isolation & purification; D015247:Deoxyribonuclease HindIII; D004587:Electrophoresis, Agar Gel / Q000295:instrumentation / Q000379:methods*; D008970:Molecular Weight; D016133:Polymerase Chain Reaction
D004279:DNA, Viral; D004247:DNA; D015247:Deoxyribonuclease HindIII