DGGE Denaturing Gradient Gel Electrophoresis System

Denaturing Gradient Gel Electrophoresis (DGGE) is a powerful genetic analysis technique that can be used for detecting single base changes and polymorphisms in genomic, cloned, and PCR amplified DNA. Two of the most valuable uses for DGGE in human genetics are in directly detecting single base changes that cause disease and in detecting polymorphisms with DNA probes for genetic-linkage analysis. 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.

In collaboration with two of the technique originators C.B.S. SCIENTIFIC has designed five different sized DGGE Systems which are reliable and easy-to-use. The DGGE-1001 is a smaller 2 gel system and features one dual gel cassette. The DGGE-2001 has a larger buffer tank and features two single gel cassettes. The DGGE-2401 has the same buffer tank as the DGGE- 2001 but features two dual gel cassettes, for a capacity of 4 gels per run. Also available is the DGGE-4001 which has a larger buffer tank and four single cassettes. Finally there is the DGGE-4801 which features the larger buffer tank and four dual cassettes enabling the researcher to run up to 8 gels simultaneously. Improvements to the technique include: a simplified method for casting perpendicular and vertical gels using Gel Wrap®, single or dual gel cassettes have been redesigned eliminating the use of agarose plugs, the need for an external peristaltic pump for buffer cycling has been eliminated, bulky screw clamps have been replaced with polypropylene spring clamps, buffer tank dimensions have been changed to use bench space more efficiently, and a safety cover with an electrical interlock has been added for protection and to help maintain temperature and reduce evaporation.

Results

Ethidium Bromide stained DGGE gel of B- Globin gene (from S.E. Asian population) after PCR of fragment II, N= Normal allele, E = mutation on codon 26, 17 = mutation on codon 17, Patient Control (IVS-1) = intervening sequence. 1. Samples run on a 45-75% gradient for 5 hours at 60 C at 200 V constant current in TAE.