PCR analysis was also carried using VirD gene-specific primers to check the contamination of hairy roots with residual Agrobacterium.
A 2.7-µL aliquot of each processed sample was amplified by PCR using the human β-globin GH20/PC04 primers to check DNA integrity [51] and two HPV generic primer sets were employed for detection, to prevent underestimation in the determination of viral prevalence caused by the use of a single HPV identification set [52].
Successful incorporation of the mutations was confirmed by DNA sequence analyses, using forward and reverse primers to check the entire AQP1 sequence, and confirm the absence of random mutations.
We measured DNA concentrations of 15 hordein clones as described previously in DNA, RNA and cDNA section and carried out the qRT-PCR reactions with selected primers to check validation of the system.
Genomic DNA of P. propionicicum (DSM 15578), S. fumaroxidans (DSM 10017), S. pfennigii (DSM 10092), S. wolinii (DSM 2805M), S. sulfatireducens (DSM 16706) and T. acetatoxydans (isolated at the Dept. of Microbiology, Swedish University of Agricultural Sciences) was subjected to PCR with the pct primers to check possible product formation.
White colonies were grown in 100 μL LB (+ampicillin, 75 mg L−1) in 96-well plates (IBL, Austria) at 37 °C, 180 rpm, for 6 h. 1 μL culture was used for colony PCR using M13 primers to check for presence and length of inserts.
A negative control (no-template control) was included in each primer assay to check for the formation of primer-dimers.
Nontemplate controls were included for each primer pair to check for significant levels of any contaminants.
Non-template controls were included for each primer pair to check for significant levels of any contaminants.
As recommended by Vekemans et al. [ 37], the correlation between AFLP band size and frequency among samples was assessed for each primer combination to check for potential homoplasy.
