Using the same RNA samples (Col-0; mock and TMV-Cg-infected, 3dpi) as those used for small RNA sequencing, 5' RACE products were detected for only two of the 16 predicted targets tested.
The SBS library SPLW was constructed using the same RNA used to construct the PLW MPSS library.
The expression patterns of twelve candidate genes were validated using the same RNA samples as in the microarray analysis.
To validate our MPSS results, we made and sequenced an SBS library using the same RNA sample that was used for the PLW library.
However, it has lower sensitivity compared with NASBA-ELISA, indicating that the amplification efficiency of PCR is lower than NASBA as they all using the same RNA template.
Five genes (comp9755_c0, comp11967_c0, comp29815_c0, comp36138_c0 and comp72325_c0) which have been identified may be related to drought stress response were validated by quantitative real-time PCR (qRT-PCR) using the same RNA samples as in the DGE library construction.
This analysis was performed in triplicate using the same RNA samples as for microarray analysis.
Real time PCR reactions were carried out using the same RNA samples, which were used for microarrays as described earlier.
As a point of reference, technical replicates using the same RNA samples performed at 2 3 months apart have a correlation coefficient of 0.96 0.98.
A total of 27 genes with different levels of expression detected using microarray were selected for real-time PCR verification using the same RNA purified for microarray hybridizations.
We selected testes and ovaries for in situ hybridization analysis using the same RNA probes that were used in our Northern blots, but this time labeled with DIG (see Materials and Methods).
