We describe the synthesis of select nucleoside analog phosphoramidites and their incorporation into RNAs that mimic known editing sites by solid phase synthesis, and analyze the interaction of these synthetic RNAs with ADARs using deamination kinetics and quantitative gel mobility shift assays.
These RNAs were added to a liver polyA sample prior to cDNA synthesis and analyzed as part of the overall experiment.
RNA was extracted, subjected to cDNA synthesis and analyzed in qPCR.
Exponentially growing MCF7 cells transfected with p27wt, p27P69L or p27W76X were incubated with Cycloheximide (CHX) to block new protein synthesis and analyzed at different time points thereafter.
As an alternative approach, cells were synchronized and treated at various times prior to the onset of DNA synthesis and analyzed for survival and/or the frequency of mutations.
(12) All of the ChM-I mimetic peptides used in this study were synthesized by standard Fmoc solid phase peptide synthesis, and analyzed by reverse-phase HPLC and mass spectrometry (Fig. 2).
This chapter discusses a new tool that is capable of reducing the complexity of the process synthesis problem and analyzing a tradeoff among the environmental impact, economy, and robustness of the process.
We proposed a new tool that is capable of reducing the complexity of the process synthesis problem and analyzing a trade-off between the environmental impact, economy and robustness of a chemical process.
One way to approach this is to employ suitable representations of the protein synthesis process and analyze this in silico.
A popular and low cost method of mutation detection has been the protein truncation test (PTT), where target regions of BRCA1/2 are PCR amplified, transcribed/translated in a cell-free protein synthesis system and analyzed for truncated polypeptides by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and autoradiography.
