The 18S rDNA PCR amplicons were then separated by DGGE
choosing a gradient between 30% and 55% (urea and formamide) and 9% polyacrylamide and running conditions of 80 V for 18 h in 1XTris Acetate EDTA buffer (TAE) (10X TAE: 48.4 g Tris, 11.4 mL Acetic Acid, 3.7 g EDTA, distilled water to 1000 mL) at 60[degrees]C (for more details about the procedure, see Yan et al., 2007; Wu et al., 2009).
Para las corridas de los geles DGGE
se determino que a las 20 horas, a 70 V y 60 [grados]C se obtienen bandas con la separacion y resolucion adecuada para la interpretacion de los geles.
of the amplified gene sequences was carried out using a DCode System (universal mutation detection system; Bio-Rad).
The similarity dendrograms of bacterial 16S DGGE
fingerprints from peach orchard showed that bacterial genetic diversity in the sites under drip emitters (PIW and PCW) were discriminated from patterns of sites along the inter-rows (PID and PCD), with Pearson similarity coefficients ranging from 94.2 to 98.7 for 16S rDNA (Fig.
Here we focus on two common DNA- and RNA-based fingerprinting methods, DGGE
and TRFLP analysis.
Until recently, our mutation detection strategy consisted of direct sequencing of the large exon 11 of both BRCA1 and BRCA2 and DGGE
for all other coding exons (3, 4).
Denaturing gradient gel electrophoresis (DGGE
) is a powerful technique for separation of double-stranded DNA molecules of nearly identical size according to their nucleotide composition.
Separation of the amplified bacterial community DNA via DGGE
was performed using the Bio-Rad D GENE system (Bio-Rad, Hercules, CA) with slight modifications to the manufacturer's instructions.
Recently, utilization of PCR primers with GC clamps and subsequent analysis by DGGE
has been shown to significantly improve the PCR analysis of T-cell clonality.[6,7,16] Assays based on the detection of PCR products of TCR genes by DGGE
require preparation of a gradient gel by a special instrument.
Nishino et al  suggested an association between the appearance of Enterobacter and high acetic acid content in tropical guinea grass silages by denaturing gradient gel electrophoresis (DGGE
) analysis, however, the abundance of this genus was unclear, as DGGE
analysis is not quantitative.
Soil contaminated with petroleum and undergoing remediation was found enriched significantly for Methanosarcinales strains with a denaturing gradient gel electrophoresis (DGGE
) method .