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Riboprobes containing Dig-11-UTP were synthesized with T3 or T7 polymerase (Promega) from PCR product templates.
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The amplification mix comprised 19 µl of mix and 1 µl of PCR product template from the first amplification round.
Following this, 18 μL reactions were prepared containing 20 ng PCR product template and 4 pmol M13 primer (Biomers).
After further elongations of the advanced starter unit without reduction, the product template (PT) domain of the nrPKS directs first ring closure by regiospecific aldol condensation.
The sequencing reaction required 2 μl of Premix, 5 pmol of sequencing primer and 0.2 μg of the PCR product template in a total volume of 10 μl.
However, the built-in product template (PT) domains of PKS4 cyclized most of the products in a fungal-specific regioselectivity, which is considerably different than those of type II PKS products (Thomas 2001).
However, since only five samples were studied it was not possible to evaluate accurately the efficacy of BeadArray on MDA product template, including estimation of sample exclusion and failure rates.
Like CCIG_05377, GME5066_g contains a starter unit acyl-carrier protein transacylase (SAT), ketosynthase (KS), acyltransferase (AT), product template (PT), two acyl-carrier proteins (ACPs) and a thioesterase (TE) domain.
Megasynthases include a starter-unit acyltransferase (SAT), a KS, a malonyl-CoA:acyl carrier protein (ACP) transacylase (MAT), a product template (PT), an acyl-carrier protein (ACP), and a thioesterase/claisen-cyclase (TE/CLC) (see [ 24] and the references therein).
These diverse natural product template sequences are flanked by highly conserved cleavage sites that ultimately direct the excision and macrocylization of the mature cyanobactin from the precursor peptide [ 25– 27].
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CEO of Professional Science Editing for Scientists @ prosciediting.com