The transcript levels for the MYB genes were measured in all five lines, myb28-1, myb29-2, myb29-2, myb76-1 and myb76-2, to determine whether the T-DNA insertions resulted in a loss of transcript.
The pathogenic mt-tRNAval mutation, which causes a loss of transcript stability when it is not charged with the correct amino acid, can be suppressed by the full length leucyl tRNA synthetase (LARS2) but not by alanyl, or phenylalanyl tRNA synthetases.
In order to assess whether repression by Pum-Brat leads to a loss of mRNA transcripts, we initially measured GFP-mad3′ transcript levels in S2 cells, with and without Pum-Brat co-transfection.
A loss of Sox2 transcript in day 4 Sox2-siRNA morulae (Figures 4A and S4) was observed compared to incubator-control (untreated) morulae, in which Sox2 transcripts were always detected.
To verify that the insertion of the selectable marker cassette resulted in a loss of the transcript for the pmt1 and pmt 4 mutants we performed real time PCRs with PMT1, PMT2, and PMT4 specific primers on cDNA derived from the serotype D mutants grown at 30°C.
Reduced levels of plastid 16S and/or 5S rRNAs in mutants of group I imply a severe drop in translation rates and consequently a general loss of transcripts synthesized by the PEP.
This mutation has been shown to result in aberrant splicing and a loss of the TMEM70 transcript [ 8].
Determination of ELP3 expression in mop3-1 demonstrated a loss of full-length transcript; however, when analyzing the genomic ELP3 locus in mop3-1, we could not characterize a T-DNA insertion but found evidence for rearrangements at this chromosomal position (Leitner, 2011).
Moreover, biallelic loss of the gene or transcriptional silencing of p53, both of which have been reported in OSCC cell lines, results in a complete loss of transcript in tumour cells (Hauser et al, 2002).
A cuvette analysis of emission profiling of different RNAs with PY is unattainable due to difficulties associated with the extraction and isolation of RNA subtypes where a loss of up to 90% of transcripts can occur [ 5].
