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Operating instructions – Hoefer HE-PLUS System User Manual

Page 11

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Operating Instructions

A. Guidelines for Selecting Electrophoresis 
Buffers and Gel Concentrations

The two most commonly used buffers for hori-
zontal electrophoresis of double stranded DNA
in agarose gels are Tris-Acetate-EDTA (TAE)
and Tris-Borate-EDTA (TBE). While the resolv-
ing powers of these buffers are very similar,
the relative buffer capacities are very different,
conferring different run attributes which are
summarized below:

TAE

Tris-acetate has traditionally been the more 
commonly used buffer. However, its relatively low 
buffer capacity will become exhausted during 
extended electrophoresis, making buffer recirculation 
necessary in runs exceeding 140 mA-hours. Potential 
advantages of using TAE buffer over TBE buffer 
include superior resolution of supercoiled DNA 
and approximately 10% faster migration of double-
stranded linear DNA fragments.

TBE

Tris-borate’s significantly greater buffering capacity 
and its relatively low current draw eliminates the 
need for recirculation in all but the most extended 
runs ( > 300 mA-hours). TBE buffer systems are not 
recommended when fragments are to be recovered 
from the gel after electrophoresis.