PMID- 9895335
OWN - NLM
STAT- MEDLINE
DCOM- 19990128
LR  - 20220129
IS  - 0009-9147 (Print)
IS  - 0009-9147 (Linking)
VI  - 45
IP  - 1
DP  - 1999 Jan
TI  - Validation of double gradient denaturing gradient gel electrophoresis through 
      multigenic retrospective analysis.
PG  - 35-40
AB  - Among established techniques for the identification of either known or new 
      mutations, denaturing gradient gel electrophoresis (DGGE) is one of the most 
      effective. However, conventional DGGE is affected by major drawbacks that limit 
      its routine application: the different denaturant gradient ranges and migration 
      times required for different DNA fragments. We developed a modified version of 
      DGGE for high-throughput mutational analysis, double gradient DGGE (DG-DGGE), by 
      superimposing a porous gradient over the denaturant gradient, which maintains the 
      zone-sharpening effect even during lengthy analyses. Because of this innovation, 
      DG-DGGE achieves the double goals of retaining full effectiveness in the 
      detection of mutations while allowing identical run time conditions for all 
      fragments analyzed. Here we use retrospective analysis of a large number of 
      well-characterized mutations and polymorphisms, spanning all predicted melting 
      domains and the whole genomic sequence of three different genes--the cystic 
      fibrosis transmembrane conductance regulator (CFTR), the beta-globin, and the p53 
      genes--to demonstrate that DG-DGGE may be applied to the rapid scanning of any 
      sequence variation.
FAU - Cremonesi, L
AU  - Cremonesi L
AD  - Instituto di Ricovero e Cura a Caraterre Scientifico, O San Raffaele, Unita di 
      Genetica e Diagnostica Molecolare, Via Olgettina 60, 20132 Milan, 
      Italy.cremonisi@laura.hsr.it
FAU - Carrera, P
AU  - Carrera P
FAU - Fumagalli, A
AU  - Fumagalli A
FAU - Lucchiari, S
AU  - Lucchiari S
FAU - Cardillo, E
AU  - Cardillo E
FAU - Ferrari, M
AU  - Ferrari M
FAU - Righetti, S C
AU  - Righetti SC
FAU - Zunino, F
AU  - Zunino F
FAU - Righetti, P G
AU  - Righetti PG
FAU - Gelfi, C
AU  - Gelfi C
LA  - eng
GR  - E.0555/TI_/Telethon/Italy
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Clin Chem
JT  - Clinical chemistry
JID - 9421549
RN  - 0 (CFTR protein, human)
RN  - 0 (Tumor Suppressor Protein p53)
RN  - 126880-72-6 (Cystic Fibrosis Transmembrane Conductance Regulator)
RN  - 9004-22-2 (Globins)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Cystic Fibrosis Transmembrane Conductance Regulator/genetics
MH  - DNA/*genetics
MH  - DNA Mutational Analysis/*methods
MH  - Electrophoresis/*methods
MH  - Globins/genetics
MH  - Humans
MH  - Mutation
MH  - *Nucleic Acid Denaturation
MH  - Polymorphism, Genetic
MH  - Reproducibility of Results
MH  - Tumor Suppressor Protein p53/genetics
EDAT- 1999/01/23 00:00
MHDA- 1999/01/23 00:01
CRDT- 1999/01/23 00:00
PHST- 1999/01/23 00:00 [pubmed]
PHST- 1999/01/23 00:01 [medline]
PHST- 1999/01/23 00:00 [entrez]
PST - ppublish
SO  - Clin Chem. 1999 Jan;45(1):35-40.