White colonies were further grown in LB broth medium with ampicillin, plasmid was isolated and subjected to restriction analysis
using EcoRI and SalI.
Amplified Ribosomal DNA Restriction Analysis
(ARDRA) is the extension of the RFLP (restriction fragment length polymorphism) technique encoding the small (16s) ribosomal subunit of bacteria.
2013) and restriction analysis
of chloroplast fragments (Parfitt and Badenes, 1997) in order to resolve the phylogenetic relationships.
Three restriction enzymes ( Hha I , BstU I, and Mbo I) were used in restriction analysis
The objectives of this study were to investigate the stem and root nodulation associated with these species and to perform phenotypic and molecular characterization, in the latter case by restriction analysis
of the 16S rDNA gene, of bacterial isolates obtained from nodules of species of the Discolobium and Aeschynomene genera in the Pantanal.
The isolates from lungs, kidney, heart, intestine, liver, and bone marrow all harbored the same virulence-associated factors (iucD, colV, iss, mat, fimC, ompA, traT crl, csgA vgrG, and hcp), yielded the same band pattern in amplified ribosomal DNA restriction analysis
, and were allocated to the E coli Reference Collection group B1.
The restriction analysis
by PstI of the positive PCR samples further subcategorized into two genotypes (Fig.
Amplified fragments were confirmed by restriction analysis
and DNA sequencing.
we compared nucleotide sequence identity to reference strains and carried out multilocus analysis using ompA-ompB sequences and in silico PstI and RsaI restriction analysis
of ompA sequences (7).
Detection of Recombinants: To analyze the presence and orientation of the DNA insert into recombinant clones, some methods including colony PCR, restriction analysis
and sequencing were performed depending on the purpose.
We will use hMeDIP-sequencing and combined glycosylation restriction analysis
to measure the differential levels of 5-methylcytosine and 5-hydroxymethylcytosine marks.
The final construct, pGEM-ura3, was confirmed by restriction analysis