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- Denaturing Gradient Gel Electrophoresis (DGGE) | SpringerLink
Denaturing gradient gel electrophoresis (DGGE) is a robust method for point mutation detection that has been widely used for many years () It is a polymerase chain reaction (PCR)-based method, the principle being the altered denaturing temperature of a PCR product with a mutation compared to the wild-type product (see Chapter 6)
- Denaturing Gradient Gel Electrophoresis - an overview . . .
TDGS is based on denaturing gradient gel electrophoresis (DGGE) as the mutation detection principle, in combination with PCR amplification to prepare the target sequences In DGGE, DNA fragments are subjected to electrophoresis in a polyacrylamide gel against a gradient of ever higher temperature or chemical denaturants (i e a mixture of urea
- Denaturing Gradient Gel Electrophoresis (DGGE) as a Powerful . . .
Optimizing DGGE conditions Three pivotal conditions needed to be optimized for accurate resolution: proportion of urea formamide as key denaturant components; acrylamide percentage as the molecular weight separation parameter and lastly, fine-tuning the primer to substrate ratio to avoid intermediate artifacts known to occur during the amplification step due to the complexity of the added GC
- mutation analysis - Bio-Rad
in the mutant samples The ability to resolve the heteroduplex fragments and the mutant homoduplex fragment in the mutant samples makes it possible to distinguish between samples that are mutant or wild type From the results, it was possible to detect single-base substitutions in the mutant samples from the wild-type sample using parallel DGGE
- Mutation detection by denaturing gradient gel electrophoresis . . .
Denaturing gradient gel electrophoresis (DGGE) allows the rapid screening for single base changes in enzymatically amplified DNA The technique is based on the migration of
- Improvements in gel composition and electrophoretic . . .
Denaturing gradient gel electrophoresis (DGGE) is believed to be the most powerful pre-screening method for mutation detection currently available, being used mostly on an exon-by-exon basis Broad-range DGGE for the analysis of multiple fragments or an entire gene is rarely applied
- Constant Denaturant Gel Electrophoresis (CDGE): Anne-Lise . . .
The optimal conditions for the CDGEs are determined using perpendicular DGGE The PCR product from a wild type sample and the mutant control are mixed 1:1, heated for 3 minutes at 94oC and reannealed at 65oC for 1 hour to allow formation of heteroduplexes before loading into the long well along the top of the gel
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