The discrepancy could be due to the differences in the methodologies of transcripts measurements; while Ishihama et al. used both semi-quantitative endpoind RT-PCR and in situ hybridisation [ 32], in this study we used qPCR which is regarded as a gold standard for the quantification of nucleic acids [ 33].
To test the efficiency of this ISH methodology, localization of transcripts of mEMK, a mouse homologue of Nematoda par-1 [45], [45], was also examined on the oocytes and zygotes (Figure S1E).
The assembly procedure may include already available reference sequences (guided approaches), as in the case of transcript assemblies based on a genome reference, or they are based on reference-free methodologies (de novo) [77].
Figure 2 shows a very high correlation (Pearson r-value of 0.7757 at a p-value <0.0001) for transcripts detected by both methodologies, indicating that our approach of estimating transcript abundance from RNA-seq data is accurate over a wide range of transcript expression levels.
End synopsis of transcript.
Appendix D. Sample of transcript coding.
This methodology provides a catalogue of transcripts that are actively being translated.
Existing guidelines, define an optimum response to therapy by the attainment of MMR by 18 months of the start of therapy [ 11, 19], with current methodology expressing transcript numbers per microgram of leukocyte RNA [ 20] or as a ratio of BCR-ABL1/ABL1 on a log scale [ 21, 22].
Furthermore, the bias in methodologies of RNA-seq and proteomics can result in the discordance between detected transcripts and proteins.
Differences in the detection of transcripts depending upon methodology suggests that not all transcripts are treated equally during sample preparation, conversion into cDNA, amplification, and other steps.
