PMID- 8922636
OWN - NLM
STAT- MEDLINE
DCOM- 19970220
LR  - 20220129
IS  - 0736-6205 (Print)
IS  - 0736-6205 (Linking)
VI  - 21
IP  - 5
DP  - 1996 Nov
TI  - Temperature-programmed capillary electrophoresis for detection of DNA point 
      mutations.
PG  - 926-28, 930, 932
AB  - A method for unambiguous determination of point mutations in genomic DNA, based 
      on electrophoresis in thin capillaries, is reported here. The method is based on 
      the principle of temperature gradient gel electrophoresis (TGGE), a variant of 
      denaturing gradient gel electrophoresis (DGGE), and exploits the differential 
      melting of mutant and wild-type PCR-amplified DNA fragments during 
      electrophoresis through a temperature gradient. Unlike TGGE, where the 
      temperature gradient exists along the separation space, the denaturing 
      temperature gradient in the fused-silica capillaries is time-programmed, so as to 
      reach the melting points (Tms) of all species under analysis prior to 
      electrophoretic transport past the detector window. The DNA fragments are 
      injected in a capillary maintained (by combined chemical and thermal means) just 
      below the expected Tm values. The temperature increment applied is typically 
      minute (1 degree -1.5 degrees C) and the sweep speed is rather shallow (e.g., 
      0.05 degree C/min). Additionally, the denaturing thermal gradient is not 
      controlled externally, but generated internally by Joule heat produced by voltage 
      ramps. Point mutants are fully resolved into a spectrum of four bands, with a 
      dynamic range extending from 45 degrees C (low melters) up to 70 degrees C for 
      high melters. The present method can thus be universally applied to any type of 
      point mutation.
FAU - Gelfi, C
AU  - Gelfi C
AD  - ITBA, CNR, Milano, Italy.
FAU - Cremonesi, L
AU  - Cremonesi L
FAU - Ferrari, M
AU  - Ferrari M
FAU - Righetti, P G
AU  - Righetti PG
LA  - eng
GR  - E.0153/TI_/Telethon/Italy
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Biotechniques
JT  - BioTechniques
JID - 8306785
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - DNA/*analysis
MH  - Electrophoresis, Capillary
MH  - *Point Mutation
MH  - Temperature
EDAT- 1996/11/01 00:00
MHDA- 1996/11/01 00:01
CRDT- 1996/11/01 00:00
PHST- 1996/11/01 00:00 [pubmed]
PHST- 1996/11/01 00:01 [medline]
PHST- 1996/11/01 00:00 [entrez]
AID - 10.2144/96215rr06 [doi]
PST - ppublish
SO  - Biotechniques. 1996 Nov;21(5):926-28, 930, 932. doi: 10.2144/96215rr06.