To test the effect of template copy number on ExCyto PCR amplification, we amplified clones from a random genomic DNA library in high, low, and single copy vectors (Figure 3).
I appreciate the idea of using genome-wide computational analysis of protein domains to test the contribution of templating mechanisms to epigenetic stability; something definitely could be done with this.
A moving stimulus was used as a template to test the effect of motion perception on subsequent arm movement execution.
We chose to test both orientations of the tag to assess its influence on the protein folding, and to test the flexibility of the developed template synthesis protocol.
Each run included a control free of template to test the assay reagents for contamination.
To overcome PCR bias in DNA methylation analysis, we optimized the annealing temperature and type of DNA polymerase: 0%, 50% and 100% methylated control DNA (Epitect methylated human control DNA; QIAGEN) was used as template to test the linearity of the protocol.
To test the integrity of the cDNA templates, a pair of control primers for the β-actin (GenBank Acc. JQ822245) of A. ipsilon was used.
To test the use of PCR product as template, the amplicons from these reactions were diluted 1 100 in ddH2O, and 2 μl of the dilution was used to amplify an internal fragment with primers TE4F and TE4R.
To test the sensitivity of this method to template concentration, we performed assays on wild-type and D2 homozygous genomic DNAs ranging in concentration from 1 to 180 ng/reaction.
