C.B.S. Scientific TTGEK-2401-220 - Manual (Page 5)

TTGE Instruction Manual, version 8/30/2011
5
www.cbsscientific.com
SECTION 1
General Information
1.1 Introduction
DESCRIPTION
The T
emporal
T
emperaTure
G
raidenT
e
lecTrophoresis
s
ysTem
allow the researcher to choose which technique is
most applicable to their investigation. Both DGGE and TTGE rely on establishing a gradient of either solvent
(Urea/Formamide) or temperature in which the target fragments will undergo conformational transition (melt). This
sequence dependent information will determine the theoretical melting behavior of the target fragment after PCR
amplification. If the sequence is known, then primer/probe design can be made using certain software design
programs (1). If not, the melting range can be revealed by running perpendicular DGGE/TTGE gels.
C.B.S. SCIENTIFIC has designed two different T
emporal
T
emperaTure
G
raidenT
e
lecTrophoresis
s
ysTems
which are
reliable and easy-to-use. The TTGEK-2001 is a 2 gel system and includes two single gel cassettes. The TTGEK-2401
is a four gel system which includes two dual gel cassettes. The systems feature: programmable heater/stirrer which
can be programmed for DGGE or TTGE, a simplified method for casting perpendicular and vertical gels using Gel
Wrap ® , single or dual gel cassettes, an internal impellor pump for buffer cycling, polypropylene spring clamps, and a
safety cover with an electrical interlock which helps maintain temperature, reduce evaporation, and protect against
shock hazard.
DGGE & CDGE
Denaturing Gradient Gel Electrophoresis (DGGE) is a powerful genetic analysis technique that can be used for
detecting single base changes and polymorphisms in genomic (2,3), cloned, and PCR amplified DNA (3,4). Two
of the most valuable uses for DGGE in human, animal or microbial genetics are in directly detecting single base
changes that cause disease and in detecting polymorphisms with DNA probes for genetic-linkage analysis. In DGGE,
conformational transitions of multiple nucleic acid complexes are induced by an increasing concentration of solvent
(Urea/Formamide) at a constant temperature. Clinical applications of DGGE include a rapid and effective method
for screening samples for genetic mutations and variants. Also, DNA fragment melting points can be determined
using perpendicular DGGE (2). In contrast to DGGE, CDGE (Constant Denaturant Gel Electrophoresis) uses a
single solvent percentage to induce partial melting of DNA fragments as they enter the gel. The disadvantage of
CDGE is that only a single melting domain can be interrogated.
TTGE
Destabilization of nucleic acid complexes can also be studied using acrylamide gels which contain a uniform solvent
concentration (Urea/Formamide), but with an increasing temperature gradient (6). Since the temperature of the
entire gel is uniformly raised over a period of time, this technique has been termed ‘TTGE’, or Temporal Temperature
Gradient Electrophoresis (7). This technique incorporates many improvements over DGGE/CDGE especially when
studying multiple melting domains (8).