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While comparing both independently produced Chrnb2−/− animals to the selected WT strain does not completely correct for allelic effects, our analysis of two independently generated mutants allows a more certain determination of transcripts affected by the lack of CHRNB2.
Real-time quantitative PCR (qPCR) is the foremost method of choice for accurate determination of transcripts amounts.
Advantages of sequencing include the direct determination of transcripts by RNA sequencing, and the fact that sequencing allows the discovery of new genes, while the genomic diversity that can be determined by microarrays is limited to the reference sequences used for array probe design.
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The combined application of multiple approaches for transcriptional profiling is likely to provide the most robust determination of transcript levels.
Herein we present, for the first time, a comprehensive view of the coupled transcriptional and translational dynamics of the P. falciparum IDC by determination of transcript abundance and architecture together with ribosomal density and positioning.
For determination of transcript concentrations, quantitative real time RT-PCR was performed in two biological replicates.
Determination of transcript abundance was performed by RT-qPCR as described above.
Most widely used is the determination of transcript level changes using DNA microarrays.
Livers were immediately removed and frozen in liquid nitrogen for subsequent determination of transcript abundance.
Critical to this annotation are the determination of transcript sequences, and an indication of when transcripts are expressed.
Quantitative polymerase chain reaction (QPCR) is a widely applied analytical method for the accurate determination of transcript abundance.
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