Positive clones were screened (restriction analysis
) by isolation of recombinant plasmid following (Thermo scientific Miniprep Kit) manufacturer recommendations and results were visualized on 1% agarose gel.
The targeted fragments (318bp and 257bp) were resolved by agarose gel electrophoresis then subjected to restriction analysis
using Bbv1, Rsa1 and Hph1 enzymes.
In order to establish the dissimilarity between strains we conducted a restriction analysis
with PacI and PmeI (Table 2).
The positive clone after restriction analysis
was utilized for DNA sequencing (Sanger et al., 1977).
pTZ57R/Basrai TLPs containing recombinant plasmid positive clones were screened by blue-white screening method, colony PCR, and restriction analysis
. The constructs containing plasmids were prepared according to Sambrook et al.
In addition various molecular characterization studies were also carried on different species, cultivars and accessions of Pistacia including RAPD (Kafkas and Perl-Treves, 2001; Kafkas and Perl- Treves, 2002; Katsiotis et al., 2003; Golan-Goldhirsh et al., 2004), AFLP (Katsiotis et al., 2003; Golan-Goldhirsh et al., 2004; Kafkas, 2006; Karimi et al., 2009), ISSR (Kafkas, 2006), SSR, SRAP (Talebi et al., 2012), IRAP (Ghaemmaghami et al., 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 primers were synthetized by Bioneer Corporation, South Korea, and all other reagents used in PCR and restriction analysis
were purchased by Fermentas, Lithuania, part of Thermo Fisher Scientific.
In the present study, culture-independent methods, random amplified polymorphic DNA (RAPD), and amplified ribosomal DNA restriction analysis
(ARDRA) were used to examine the bacterial community and dynamics of dominant bacterial species in ginseng rhizosphere soil during the growth of P.
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.