The HS2 fragment was amplified (Table 1) using the primers HS2s and HS2as (Table 2) and inserted into the BamHI and SalI restriction sites
Assembly was achieved in the pFastBac1 donor plasmid by exploiting unique restriction sites
to concatenate the entire genome from 3 sub-clones.
Both adapters were complemented with Ins, that is, inserts of 0 to 3 bases that increase the sequence variability to preserve the lasers of the sequencer from blinding by the identical bases in the restriction sites
The insertion of the KpnI restriction site
allowed the circularization of the linear amplicon.
The forward and reverse primers contained NdeI and BamHI restriction sites
, respectively (underlined).
Furthermore, DNA sequence confirmed the presence of a BsrDI restriction site
in the tprC gene, which would be absent in subspecies pertenue.
This analysis identified a restriction site
for the restriction enzyme Fnu4HI within the 15 base-pair deletion identified in specimens of D.
; however, it also matches genera within the [alpha]-, [beta], and [gamma]-proteobacteria, and Firmicutes phyla.
The analysis of nucleotide sequence of p85a insulin gene deposited at NCBI gene bank was under taken with special reference to nucleotide position 1227 with sequence CGG or CAG created a loss of Msp1 restriction site
(CJ[down arrow]CGG) which is an advantageous diagnostic tool for identifying the insulin resistance due to genetic defects of leading on to the impaired insulin mediated PI3K activity.
An intact PCR product indicates absence of Bsm1 restriction site
(B allele), whereas the presence of the BsmI site (b allele) is denoted by two or three fragments.
For [lambda] DNA sequences, we counted the number of reads mapped to each restriction site
and ignored sites with restriction fragment lengths of <30 bp (because 25 bp was used for alignment).
Thus, the Oka vaccine strain was positive for the additional SmaI restriction site
in ORF 62 (SmaI+), while all the circulating wild-type VZV strains in India were negative (SmaI-).