All sub-cloning procedures were confirmed by using restriction enzymes and DNA sequence analysis.
The LDH gene with the His tag was subcloned into pET14b plasmids from Novagen using restriction enzymes NcoI and BamHI with standard molecular biology procedures.
Genomic DNA methylation analysis using restriction enzymes.
Participants were randomly assigned following simple 1 1 randomisation procedures using opaque and sealed envelopes to one of the two study arms (interventional or control arm) without any restriction.
We used restriction enzymes and direct sequencing.
In case no valid primers can be found around any restriction site, a fall-back mechanism will repeat this enumeration procedure using the 'next best restriction site' if present.
In the latter plasmid, the promoter upstream region can be easily substituted by a one-step cloning procedure using the unique EcoRI/KpnI restriction sites.
The present paper slightly modifies this procedure, using sca j f and sra i f as column and row restrictions in a RAS algorithm.
Standard procedures were used for restriction enzyme mediated DNA digestion, ligation, and transformation [ 29].
Standard procedures were used for restriction endonuclease digestions, electrophoresis and transfer to nylon membranes.
