Whole-genome amplification is a crucial first step in nearly all single-cell genomic analyses, with the following steps focused on its products. Bias and variance caused by the whole-genome amplification process add numerous challenges to the world of single-cell genomics. Short tandem repeats are sensitive genomic markers used widely in population genetics, forensics, and retrospective lineage tracing.
View Article and Find Full Text PDFShort tandem repeats (STRs) are polymorphic genomic loci valuable for various applications such as research, diagnostics and forensics. However, their polymorphic nature also introduces noise during in vitro amplification, making them difficult to analyze. Although it is possible to overcome stutter noise by using amplification-free library preparation, such protocols are presently incompatible with single cell analysis and with targeted-enrichment protocols.
View Article and Find Full Text PDFDeducing generic causal relations between RNA transcript features and protein expression profiles from endogenous gene expression data remains a major unsolved problem in biology. The analysis of gene expression from heterologous genes contributes significantly to solving this problem, but has been heavily biased toward the study of the effect of 5' transcript regions and to prokaryotes. Here, we employ a synthetic biology driven approach that systematically differentiates the effect of different regions of the transcript on gene expression up to 240 nucleotides into the ORF.
View Article and Find Full Text PDFAccurate and efficient gene expression requires that protein translation initiates from mRNA transcripts with high fidelity. At the same time, indiscriminate initiation of translation from multiple ATG start-sites per transcript has been demonstrated, raising fundamental questions regarding the rate and rationale governing alternative translation initiation. We devised a sensitive fluorescent reporter assay for monitoring alternative translation initiation.
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