A novel chrome-free surface pretreatment and primer combined in one-step application for aircraft coating is described.
Using the novel 5′-modified primer combined with indirect labeling method, cDNA probes can be prepared with much less total RNA (5 μg or less) without amplification producing optimal results after hybridization of arrays.
The insert orientation was confirmed by PCR using M13 forward primer combined with MutLR primer, and complementation of PAO1ΔmutL phenotype shown using the rifampicin resistance method.
In a first PCR, a specific 16S rDNA primer for Microcystis (CH) described by Rudi et al. [55] was used as forward primer combined with the universal reverse 23S rDNA primer ULR [54].
A longer full-length cDNA amplicon used the same 3' UTR reverse primer combined with a forward primer corresponding to the trans-spliced leader sequence to amplify only spliced full-length versions of the actin gene.
Briefly, RNA was reverse transcribed using a random primer combined with an oligo dT primer.
RNA was quantified, its quality verified and cDNA synthesized using an anchored oligo-T18 primer combined with random hexamers [ 57].
FAM-MGB-probe AACCGAGGCACGGACAAC was used with universal reverse primer combined with alternative forward oligos specific for either 1a isoform or 1b isoform.
The Z-PRIME Plus treatment combined with air abrasion produced the highest bond strength, followed by Z-PRIME Plus application, Metal/Zirconia Primer combined with air abrasion, air abrasion alone, and, lastly, Metal/Zirconia Primer application.
cDNA synthesis was undertaken with 5 μg of isolated total RNA per sample using MMLV Superscript III reverse transcriptase (Invitrogen) and an anchored oligo-T18 primer combined with random hexamers [ 9].
Similarly, a first reverse target-specific primer (a) and one kind of the degenerated random tagging primer (DRT) mix (A-D) were applied, in a first step, followed by two semi-nested PCR rounds using target-specific primers (b and c), that are each time nested to the previous reverse target-specific primer, combined to universal tagging primers (UAP-N1 and UAP-N2) [ 14].
