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To ensure that there is no influence on the absorbance caused by the substrates or the enzyme preparations, control reactions of all possible combinations with DCPIP were performed and no decrease in absorbance was observed (entries 1 5).
For each of the 3 cell preparations (control, G9a-shRNA or SUV39H1-shRNA treated cells), 100 nuclei were scored by two independent observers.
After measuring Ca2+transients in normal superfusion solution (HBSS), the same preparations (control, scramble and siRNA M2) were superfused with an HBSS solution containing TC 0.2 mmol/L and Ca2+ transients were recorded again after 20 min perfusion in the same conditions.
In all preparations, control samples of haploid yeast containing the MDN1 wt) gene were processed identically in order to confirm the specificity of antibody staining.
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However, cross-experimental conclusions regarding expression levels are difficult to draw due to different sample preparations, controls, etc.
It is difficult to compare our study with previous reports because most of those studies used different antigen preparations, controls and methodology [11] [19].
Still, most of these methods are in the development stage, and problems are often solved through the stability of the nanoparticle preparation, control of the crystal growth, and aggregation of the particles[11].
The inclusion of an extraction/PCR control DNA to indicate successful DNA extraction and removal of PCR inhibitors should be easily adaptable as a sample preparation control for other acellular sample types.
A drop of each exosome preparation (control, M. smegmatis and M. avium) was placed on parafilm and directly covered by a 300 mesh copper grid.
Negative controls included a reagent control (sterile water served as PCR template) and a sample preparation control (sterile water used in place of the original sample and exposed to the entire extraction protocol).
In parallel with Trx-CFP rAAV stock preparation, control CFP rAAV stock was produced by co-transfection of pAAV-CFP reporter vector, pAAV-RC and pHelper into AAV-293 cells, as described above.
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