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The absence of the DNA in the enzyme-DNA co-crystal structure, however, prevented us from examining the protein-DNA interactions through which DraIII recognizes the canonical 9-bp sequence with a three base gap in the middle.
The Molecular Inversion Probe (MIP) [14], a more advanced version of the padlock probe, contained also i) a molecular barcode for downstream chip detection, ii) a single base gap that enables single nucleotide polymorphism (SNP) detection, and iii) an exponential amplification strategy based on probe inversion and universal primer PCR amplification.
The Connector Inversion Probe (CIPer) reaction (Figure 1) shares some basic design features with a recently described assay, the PathogenMip assay [17], which includes: i) probe hybridization, single "G" base gap fill and probe circularization, ii) exonuclease based circular DNA selection, iii) probe inversion by re-linearization, and iv) universal PCR amplification.
Blood gas analysis of the groups revealed increase in the acidosis and base gap in the S/C and S/C + EP groups versus the sham groups.
A probe was retained if at least 28 of the bases matched a region in the genome with less than a two base gap within the sequence.
The 20 base gap was consistently found to be vulnerable, whether it was at the 3′ end or in the middle of the DNA strand.
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At this step, we segregated any sequences containing a frame shift (single or double base gaps or insertions) within the V3 loop.
A 6 bp run of Ts within 50 bp of the start is expanded to include all Ts with any number of single base gaps.
Such individuals review functional and technical specifications with an eye for clinical impact, potential functionality conflicts, and knowledge base gaps.
The SNP is located at this single-base gap.
The single-base gap is filled by a polymerization/ligation reaction.
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