Modifications included increasing the amount of starting mRNA to 20 μg, the addition of T4gene32 protein during first strand synthesis [ 82], the use of the plasmid pDNR-Lib (Clontech, Mountain View, CA, USA) for cDNA cloning, and the omission of radioactive labelling to monitor first strand synthesis.
For labeling, omission of the primary antibody served as a negative control.
For example, it was agreed that if a prescriber specified an indication on the prescription (such as omeprazole 'for oesophagitis'), then omission of this indication from the dispensing label was a labelling error.
Controls for specificity of immunohistochemical labeling included omission of the primary antibody and use of preimmune normal mouse and rabbit serum as previously described [26], [52] [54].
In addition, omission of the incubation step with labeled lectin from routine processing enabled us to exclude lectin-independent signal generation.
Specificity of labelling and absence of signal crossover was excluded in all cases by examination of single labelled control samples, including labelling controls with omission of primary antibody.
Control labelling was performed with omission of the first antibodies to ensure that there was no unspecific labelling of cells.
The specificity of the labeling was controlled by omission of the primary antibody.
Negative controls, which included the omission of the primary antibodies, revealed no significant labelling.
Labeling was abolished after omission of primary antibodies, ruling out nonselective binding of the secondary antibody.
Controls using omission of primary or secondary antibodies revealed no labeling.
