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Numerous studies have used viral sequencing to evaluate HIV-1 transmission linkage, but our analysis represents the first use of viral sequencing for HIV-1 transmission linkage as an integral component in the primary efficacy analysis of a large randomized HIV-1 prevention trial.
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Accordingly, we have worked to develop a low bias method that, to our knowledge, for the first time does not entail the use of viral sequence specific primers and enables next generation sequencing of the full HIV genome from a clinically relevant sample.
One technique used to reduce the chance clustering of viral sequences is the inclusion of 'local controls' - that is, viral sequences from infected individuals in the same location who are not believed to be part of the outbreak under investigation.
Resequencing DNA microarray (RMA) technology uses probes designed to identify a panel of viral sequences.
In this study both viral RNA from plasma and cell-associated proviral DNA were used as sources of templates for amplification of viral sequences.
Furthermore, these two strains were not selected a priori for favorable match of viral sequences with oligonucleotide primers used for target gene sequence amplification.
Using next-generation sequencing, the authors observe an enrichment of viral sequences in the precipitated RNA, and in particular of antisense-transcripts from the EMCV L-region.
This work is the first to estimate intrasubtype recombination of viral sequences of CRF01_AE at the host population level using NFLG sequences around the world.
Amplification of viral sequences was done in a 0.5-ml Eppendorf tube using an immunocapture PCR procedure described previously [ 16].
They took scores of viral sequences and lined them up to see what they had in common.
The isolation of substantial numbers of viral sequence variants at highly variable viral protein domains remains a major challenge.
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