Improvements in Gene Panel and Whole Exome sequencing Molecular barcoding refers to the attachment of a unique molecular sequence (eg. ATGCCA) to each individual DNA fragment in a sample. This ensures that in downstream analysis, unique sequence reads can be separated from those originating as a result of PCR duplication. A common practice regarding these PCR duplicates is to search…
Transcription profiles give accurate images of the biological processes that influence cell fates. However, library construction can introduce bias at multiple steps. For example, PCR amplification increases the number of cDNA molecules to an amount sufficient for sequencing, but also stochastically introduces errors and amplifies different molecules with unequal probabilities. The sequencing reads that have been created from the same…
If you start analyzing your gene-expression data set, many questions come to mind that can seriously affect the outcome. What if any of the PCR steps during library prep (Figure 1) has introduced an error? It might have led to a base change, which I could incorrectly score as a mutation. Did a PCR-bias favor amplification of some of the…
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