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Titolo:
COMPARISON OF BEHAVIOR OF N-SUBSTITUTED ACRYLAMIDES AND CELLULOSES ONDOUBLE-STRANDED DNA SEPARATIONS BY CAPILLARY ELECTROPHORESIS AT 25-DEGREES AND 60-DEGREES-C
Autore:
GELFI C; PEREGO M; LIBBRA F; RIGHETTI PG;
Indirizzi:
LITA,VIA FRATELLI CERVI 93 I-20090 SEGRATE MILANO ITALY CNR,IST TECNOL BIOMED AVANZATE I-20131 MILAN ITALY UNIV CALABRIA,DEPT CELL BIOL I-87036 COSENZA ITALY
Titolo Testata:
Electrophoresis
fascicolo: 8, volume: 17, anno: 1996,
pagine: 1342 - 1347
SICI:
0173-0835(1996)17:8<1342:COBONA>2.0.ZU;2-Z
Fonte:
ISI
Lingua:
ENG
Soggetto:
CROSS-LINKED POLYACRYLAMIDE; POLYMER-SOLUTIONS; FLUORESCENCE DETECTION; ZONE ELECTROPHORESIS; FRAGMENTS; MATRICES;
Keywords:
N-SUBSTITUTED ACRYLAMIDES; DNA ELECTROPHORESIS; CAPILLARY ELECTROPHORESIS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
32
Recensione:
Indirizzi per estratti:
Citazione:
C. Gelfi et al., "COMPARISON OF BEHAVIOR OF N-SUBSTITUTED ACRYLAMIDES AND CELLULOSES ONDOUBLE-STRANDED DNA SEPARATIONS BY CAPILLARY ELECTROPHORESIS AT 25-DEGREES AND 60-DEGREES-C", Electrophoresis, 17(8), 1996, pp. 1342-1347

Abstract

The behavior of N-substituted poly(acrylamides) and of cellulose in the separation of double-stranded DNA by capillary zone electrophoresis(CZE) is evaluated. Contrary to previously held belief, which attributed best separations in poly(acrylamides) to small DNA fragments (typically in the 50-1000 bp size range) and in celluloses to large DNA fragments, we show that also celluloses can achieve fine sieving of shortDNA sizes provided they are used at much higher concentrations than previously reported. For example, in the case of hydroxyethylcellulose (HEC), typically used at 0.2-0.8% concentrations, levels of 3% produceexcellent patterns, at 25 degrees C, in the 50-600 bp size range. If separations are conducted at 60 degrees C, sieving is lost in most liquid polymers. However, if the concentration of HEC is raised to 6%, and that of hydroxypropylmethylcellulose (HPMC) above 1%, sieving is fully restored. Also, N-substituted acrylamido derivatives, notably N-acryloylaminoethoxyethanol (AAEE) and N-acryloylaminopropanol (AAP) offerexcellent performance at both 25 degrees and 60 degrees C, whereas plain poly(acrylamide) irreversibly collapses at high temperatures, possibly due to intrinsic instability of the amido bond under such harsh conditions. Among all the polymers investigated, an 8% solution of poly(AAP) offers the best performance and highest theoretical plates in the 25 degrees-60 degrees C interval. Separations at high temperatures are necessary when dealing with detection of point mutations in temperature-programmed CZE and are preferred in DNA sequencing. Two additional advantages are evident when running DNA separations at 60 degrees C:a marked reduction of analysis times and a linearization of the transit times of the larger (434-587 bp) DNA fragments, in all polymers tested.

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Documento generato il 28/11/20 alle ore 12:52:58